CN108348598A

CN108348598A - The anti-C1s antibody of humanization and its application method

Info

Publication number: CN108348598A
Application number: CN201680032952.9A
Authority: CN
Inventors: 桑迪普·帕尼克; 格雷汉姆·帕里
Original assignee: Bauer Bainati American Co
Current assignee: Bauer Bainati American Co
Priority date: 2015-04-06
Filing date: 2016-04-05
Publication date: 2018-07-31
Anticipated expiration: 2036-04-05
Also published as: WO2016164358A1; PL3280440T3; SG10201909180SA; HUE061076T2; JP2021006544A; US20180092974A1; AU2022203679A1; IL254670B; US20200405852A1; JP2018513213A; JP7005483B2; SG11201707886WA; HRP20230093T1; PT3280440T; MX2017012834A; US11246926B2; CO2017011238A2; CN117285634A; KR20180006379A; US20230218753A1

Abstract

Present disclose provides the anti-C1s antibody of humanization.Present disclose provides the nucleic acid for including the nucleotide sequence for encoding the anti-C1s antibody of humanization；And include the host cell of the nucleic acid.Present disclose provides the compositions for including the anti-C1s antibody of the humanization.Present disclose provides the application methods of the anti-C1s antibody of the humanization.

Description

The anti-C1s antibody of humanization and its application method

Cross reference

Present application requires the U.S. Provisional Patent Application submitted on April 6th, 2015 No. 62/143,636 and 2015 8 The equity for the U.S. Provisional Patent Application the 62/200th, 997 that the moon is submitted on the 4th, the application case are whole by reference simultaneously Enter herein.

It introduces

Complement system is effect mechanism well known to immune response, is not only directed to pathogen and other harmful chemicals provide and prevent Shield, and injury recovery is provided.It is usually in protein existing for inactive form in vivo that complement pathway, which includes many,.Classics are mended Body approach is triggered by the activation of the first component of complement, and the first component is known as C1 compounds, is made of C1q, C1r and C1s.C1 After being combined with immune complex or other activators, the serine stretch protein of C1s components, i.e. diisopropyl fluorophosphate (DFP) (DFP)-sensitivity Enzyme, complement component C4 and C2 is to cause the activation of classic complement approach for cracking.Classic complement approach seems in many diseases and disease It works in disease.

Summary

Present disclose provides the anti-C1s antibody of humanization.Present disclose provides include the coding anti-C1s antibody of humanization The nucleic acid of nucleotide sequence；And include the host cell of the nucleic acid.Present disclose provides include the anti-C1s antibody of the humanization Composition.Present disclose provides the application methods of the anti-C1s antibody of the humanization.

Present disclose provides a kind of humanized antibodies of specific binding complement component C1s, wherein the antibody includes：a) Include the areas VH of following amino acid sequence：

(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T) SEDTAVYYCARYGYGREVFDYWGQGTTVTVSS(SEQ ID NO：26)；And b) include the areas VL of following amino acid sequence：

DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQQK(T/P)GQPPK(I/L) LIYDASNLESGIPARFSGSGSGTDFTLTISSLE(E/P)EDFA(I/V)YYCQQSNEDPWTFGGGTKVEIK(SEQ ID NO：27).In some cases, the antibody includes：A) include SEQ ID NO：10 areas VH；And b) comprising SEQ ID NO： 20 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：10 areas VH；And b) comprising SEQ ID NO：22 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：10 areas VH；And b) comprising SEQ ID NO：24 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：12 areas VH；And b) comprising SEQ ID NO：20 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：12 areas VH；And b) comprising SEQ ID NO：22 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：12 areas VH；And b) comprising SEQ ID NO：24 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：14 areas VH；And b) comprising SEQ ID NO：20 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：14 areas VH；And b) comprising SEQ ID NO：22 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：14 areas VH；And b) comprising SEQ ID NO：24 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：16 areas VH；And b) comprising SEQ ID NO：20 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：16 areas VH；And b) comprising SEQ ID NO：22 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：16 areas VH；And b) comprising SEQ ID NO：24 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：18 areas VH；And b) comprising SEQ ID NO：20 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：18 areas VH；And b) comprising SEQ ID NO：22 areas VL.In some cases, the antibody includes：A) include SEQ ID NO：18 areas VH；And b) comprising SEQ ID NO：24 areas VL.In some cases, the humanized antibody is selected from by Fab segments, F (ab ')₂Segment, scFv and Fv compositions Group.In some cases, the humanized antibody includes the heavy chain constant region of isotype IgG1, IgG2, IgG3 or IgG4.

Present disclose provides a kind of composition, it includes：A) such as above and described below humanized antibody；And b) pharmacy Upper acceptable excipient.In some cases, the composition includes tonicity agent, suspending agent, emulsifier, stabilizer, anti-corrosion It is one or more in agent, freeze drying protectant, surfactant and sugar.Present disclose provides a kind of containers, and it includes the disclosure Composition.In some cases, the container is sterile.In some cases, the container is bottle, bottle or injection Device.

Complement component cracking production in individual (for example, fluid, tissue or organ of individual) is reduced present disclose provides a kind of The horizontal method of object, the method includes by effective amount for inhibiting C1s and reducing the pyrolysis product level, to the individual Using the humanized antibody of such as above or described below disclosure, or such as the composition of above or described below disclosure. In some cases, the complement component pyrolysis product is C4 pyrolysis products (for example, C4b).In some cases, the complement Component pyrolysis product is C2 pyrolysis products (for example, C2a).In some cases, the complement component pyrolysis product cracks for C3 Product.In some cases, the individual is people.In some cases, the application is intravenous.In some cases, institute It states and applies as intramuscular.In some cases, the application is intrathecal.In some cases, the application is subcutaneous.At some In the case of, the reduction of complement component pyrolysis product level is effective to the illness for treating complement-mediated.In some cases, the benefit The illness that body mediates is isoimmunization illness.In some cases, the illness of the complement-mediated is autoimmune disorder.

Present disclose provides a kind of methods for inhibiting the complement component that C1s is mediated in individual to crack, and the method includes pressing Effectively inhibit the amount that the complement component that C1s is mediated cracks, to the people of the individual application such as above or described below disclosure Source antibody, or the composition such as above or described below disclosure.In some cases, the individual is people.At some In the case of, the application is intravenous.In some cases, the application is intramuscular.In some cases, described apply is It is intrathecal.In some cases, the application is subcutaneous.In some cases, the inhibition for the complement component cracking that C1s is mediated is to controlling The illness for treating complement-mediated is effective.In some cases, the illness of the complement-mediated is isoimmunization illness.In some cases Under, the illness of the complement-mediated is autoimmune disorder.

The method of the disease of complement-mediated or illness in individual is treated present disclose provides a kind of, the method includes by having Effect treats the disease of the complement-mediated or the amount of illness, to the people of the individual application such as above or described below disclosure Source antibody, or the composition such as above or described below disclosure.In some cases, the complement component pyrolysis product For C3 pyrolysis products.In some cases, the individual is people.In some cases, the application is intravenous.In some feelings Under condition, the application is intramuscular.In some cases, the application is intrathecal.In some cases, the application is skin Under.In some cases, the reduction of complement component pyrolysis product level is effective to the illness for treating complement-mediated.In some cases Under, the illness of the complement-mediated is isoimmunization illness.In some cases, the illness of the complement-mediated is autoimmunity Illness.

Brief description

Fig. 1 describes amino acid sequence (the SEQ ID NO of humanization VH variants 1：And the core of encoding humanized VH variants 1 10) Nucleotide sequence (SEQ ID NO：11).

Fig. 2 describes amino acid sequence (the SEQ ID NO of humanization VH variants 2：And the core of encoding humanized VH variants 2 12) Nucleotide sequence (SEQ ID NO：13).

Fig. 3 describes amino acid sequence (the SEQ ID NO of humanization VH variants 3：And the core of encoding humanized VH variants 3 14) Nucleotide sequence (SEQ ID NO：15).

Fig. 4 describes amino acid sequence (the SEQ ID NO of humanization VH variants 4：And the core of encoding humanized VH variants 4 16) Nucleotide sequence (SEQ ID NO：17).

Fig. 5 describes amino acid sequence (the SEQ ID NO of humanization VH variants 5：And the core of encoding humanized VH variants 5 18) Nucleotide sequence (SEQ ID NO：19).

Fig. 6 describes amino acid sequence (the SEQ ID NO of humanization V κ variants 1：And the core of encoding humanized V κ variants 1 20) Nucleotide sequence (SEQ ID NO：21).

Fig. 7 describes amino acid sequence (the SEQ ID NO of humanization V κ variants 2：And the core of encoding humanized V κ variants 2 22) Nucleotide sequence (SEQ ID NO：23).

Fig. 8 describes amino acid sequence (the SEQ ID NO of humanization V κ variants 5：And the core of encoding humanized V κ variants 5 24) Nucleotide sequence (SEQ ID NO：25).

Fig. 9 provides table 2, which shows the amino acid of differences between parent TNT005 VH and Exemplary humanized VH variants.

Figure 10 provides table 3, which shows the amino acid difference between parent TNT005 VL and Exemplary humanized VL variants It is different.

Figure 11 provides table 4, which shows the binding characteristic of the humanization variants of TNT005.It shows and activation C1s (" aC1s ") is bound directly and 50pM biotinylations TNT005 (" Biot-005 ") competitive bindings and the suppression to classic complement approach The data of system.

Figure 12 provides table 5, which shows the binding characteristic of the humanization variants of TNT005.Provide the humanization of TNT005 The affine data that variant is combined with people's activity C1s.

Figure 13 provides the amino acid sequence of people C1s.

Figure 14 is depicted in the pharmacokinetics of humanization TNT005 variants in the machin of (- the 43 day the 1st day) 1 phase administration (PK) feature.

Figure 15 is depicted in the medicine of humanization TNT005 variants in the machin of (- the 43 day the 32nd day) 1 phase administration for power Learn feature.

Figure 16 describes the classical pathway activity of the serum from the machin being administered (- the 43 day the 1st day) 1 phase.

Figure 17 is depicted in humanization TNT005 in the machin of 2 phases administration (the daily subcutaneous administrations of 4mg/kg continue 7 days) and becomes PK and pharmacodynamics (PD) feature of body.

Definition

Term " antibody " and " immunoglobulin " include the antibody or immunoglobulin of any isotype, holding and antigen spy It is antibody fragment (include but not limited to Fab, Fv, scFv and Fd segment) that the opposite sex combines, chimeric antibody, humanized antibody, single-stranded Antibody (scAb), single domain antibody (dAb), single domain heavy chain antibody, single domain light chain antibody, bispecific antibody, multi-specificity antibody With fusion antibody (it includes the antigen binding of antibody (herein also referred to as combining antigen) part and non-antibody proteins).Antibody can Enzyme, fluorescin etc. through (for example) radioactive isotope, generation detectable product detectably mark.Antibody can further with Other parts are conjugated, such as the member of specific binding pair, such as biotin (biotin-avidin specific binding pair Member) etc..Antibody can also be combined with solid support, including but not limited to polystyrene board or bead etc..The term is also contained Cover Fab ', Fv, F (ab ')₂And/or keep the other antibody fragments combined with antigentic specificity and monoclonal antibody.As herein Used, monoclonal antibody is the antibody generated by a kind of same cell, and whole cells are answered by individual cells by the cell repeated It makes and generates.That is, clone's object of cell only generates monospecific antibody type.Although hybridoma generation technique can be used to generate Dan Ke Grand antibody, but other generation methods well known by persons skilled in the art can also be used (for example, being originated from antibody phage display library Antibody).Antibody can be unit price or divalent.Antibody can be Ig monomers, " Y shape " molecule being made of four polypeptide chains：It is logical Cross two heavy chains and two light chains of disulfide bond connection.

" Humanized immunoglobulin " or " humanized antibody " refers to comprising separate sources as used herein, the term The immunoglobulin of immunoglobulin part, wherein at least one part include the amino acid sequence of people source.For example, humanization Antibody may include the immunoglobulin for being originated from the non-people source such as mouse with necessary specificity and the immune ball from people source The part (for example, gomphosis immunoglobulin) of protein sequence is connected to one by routine techniques (for example, synthesis) through chemical method It rises or is prepared into continuous polypeptide (for example, the DNA of the protein portion of encoding chimeric antibodies can be expressed using technique for gene engineering Generate continuous polypeptide chain).Another example of Humanized immunoglobulin is containing one or more comprising from non-people source The immunoglobulin chain of the framework region of the CDR of antibody and light chain and/or heavy chain from people source immunoglobulin (for example, With or without the CDR grafted antibody of frame variation).Term Humanized immunoglobulin is also covered by chimeric or CDR transplanting single-chain antibodies. See, e.g., Cabilly et al., U.S. Patent No. 4,816,567；Cabilly et al., European Patent No. 0,125,023 No. B1；Boss et al., U.S. Patent No. 4,816,397；Boss et al., European Patent No. 0,120,694 B1； Neuberger, M.S. et al., WO 86/01533；Neuberger, M.S. et al., European Patent No. 0,194,276 B1； Winter, U.S. Patent No. 5,225,539；Winter, European Patent No. 0,239,400 B1；Padlan, E.A. et al., European Patent Application No. 0,519,596 A1.About single-chain antibody, referring further to, Ladner et al., U.S. Patent No. 4,946, No. 778；Huston, U.S. Patent No. 5,476,786；And Bird, R.E. et al., Science, 242：423-426 (1988))。

It is, for example, possible to use the gene (for example, cDNA) that synthesis and/or recombinant nucleic acid prepare humanization chain needed for coding comes Generate Humanized immunoglobulin.It is, for example, possible to use PCR method of mutagenesis changes the DNA sequence dna of encoding human or humanization chain, such as DNA profiling from previous humanization variable region come build encoding humanized variable region nucleic acid (for example, DNA) sequence (referring to For example, Kamman, M. et al., Nucl.Acids Res., 17：5404(1989))；Sato, K. et al., Cancer Research, 53：851-856(1993)；Daugherty, B.L. et al., Nucleic Acids Res., 19 (9)：2471- 2476(1991)；And Lewis, A.P. and J.S.Crowe, Gene, 101：297-302(1991)).Using these or it is other suitable Method, can also easily produce variant.For example, the variable region that can be cloned with mutagenesis, and it is required that coding can be selected to have The sequence of the variant of specificity is (for example, come from phage library；See, for example, Krebber et al., U.S. Patent No. 5,514, No. 548；Hoogenboom et al., WO 93/06213 is published on April 1st, 1993)).

" antibody fragment " includes a part for complete antibody, for example, the antigen binding domain or variable region of complete antibody.Antibody The example of segment includes Fab, Fab ', F (ab ')₂With Fv segments；Double antibody；Linear antibodies (Zapata et al., Protein Eng.8(10)：1057-1062(1995))；Domain antibodies (dAb；Holt et al. (2003) Trends Biotechnol.21： 484)；Single-chain antibody molecules；With the multi-specificity antibody formed by antibody fragment.Papain digestion of antibodies generates two titles For " Fab " segment, respectively there is the same antigen binding fragment of single antigen binding site and remaining " Fc " segment, title Reflect the ability for being easy crystallization.Papain enzymatic treatment generates tool there are two antigen binding site and remains able to be crosslinked anti- Former F (ab ')₂Segment.

" Fv " is the minimum antibody fragment containing complete antigen recognizing and binding site.The region is by close, non-covalent The dimer of one heavy-chain variable domains of association and light variable domains composition.It is each variable in this configuration Three CDR of structural domain interact to limit V_H-V_LAntigen binding site on dimer interface.Generally speaking, six CDR Antibody is assigned with antigen-binding specificity.However, although affinity is lower than entire binding site, even if single variable domains (or only including the half Fv of the CDR of three original specificity of confrontation) can also identify and combine antigen.

First constant domain (CH of " Fab " segment also constant domain containing light chain and heavy chain₁).Fab segments with Fab ' segments are the difference is that in heavy chain CH₁The carboxyl terminal of structural domain is added to several residues, including comes from antibody hinge One or more cysteines in area.Fab '-SH are that the cysteine residues to wherein constant domain carry free sulphur herein The title of the Fab ' of alcohol groups.F(ab′)₂Antibody fragment is generated initially as pairs of Fab ' segments, has hinge between them Chain cysteine.Other chemical couplings of antibody fragment are also known.

" light chain " of antibody (immunoglobulin) from any vertebrate class can be based on the amino of its constant domain Acid sequence is assigned as one of two significantly different types (being known as κ and λ).According to the amino acid sequence of the constant domain of its heavy chain Immunoglobulin, can be assigned as different classes of by row.There are the other immunoglobulins of five major class：IgA, IgD, IgE, IgG and IgM, and several in these classifications are further divided into subclass (isotype), such as IgG1, IgG2, IgG3, IgG4, IgA And IgA2.The subclass can be further divided into the type of such as IgG2a and IgG2b.

" scFv " or " sFv " or " scFv " antibody fragment include the V of antibody_HAnd V_LStructural domain, wherein these structural domains are deposited It is in single polypeptide chain.In some embodiments, Fv polypeptides also include between V_HAnd V_LPeptide linker between structural domain, It enables sFv to form the required structure for antigen binding.Comment for sFv, referring to Pluckthun in The Pharmacology of Monoclonal Antibodies, volume 113, Rosenburg and Moore are edited, Springer- Verlag, New York, the 269-315 pages (1994).

Term " double antibody " refers to that there are two the small antibody fragments of antigen binding site for tool, and the segment is included in same polypeptide Chain (V_H-V_L) in light variable domains (V_L) connection heavy-chain variable domains (V_H).It cannot be allowed by using too short The connector matched between two structural domains on same chain, forces the complementary domain of the structural domain and another chain to match and produce Raw two antigen binding sites.In such as EP 404,097；WO 93/11161；And Hollinger et al. (1993) Proc.Natl.Acad.Sci.USA 90：Double antibody is described more fully in 6444-6448.

As used herein, term " affinity " refers to the balance that two kinds of medicaments (such as antibody and antigen) invertibity combines Constant and it is expressed as dissociation constant (K_D).Affinity can at least 1 times higher to the affinity of unrelated amino acid sequence than antibody, At least 2 times high, at least 3 times high, at least 4 times high, at least 5 times high, at least 6 times high, at least 7 times high, at least 8 times high, height at least 9 Again, at least 10 times high, at least 20 times high, at least 30 times high, at least 40 times high, at least 50 times of height, at least 60 times high, height at least 70 Again, at least 1,000 times or more of at least 80 times high, at least 90 times high, at least 100 times of height or height are again.Parent of the antibody to target protein Can be that for example, about 100 nanomoles (nM) to about 0.1nM, about 100nM to about 1 picomole (pM) or about 100nM to about 1 fly with power Mole (fM) or more.As used herein, term " affinity " refer to the compound of two or more medicaments after dilution To the resistance of dissociation.Term " immunoreactivity " and " preferential to combine " are herein for antibody and/or antigen-binding fragment It is used interchangeably.

Term " in conjunction with " refers between two molecules, due to for example covalently, electrostatic, hydrophobicity and ion and/or hydrogen bond phase Interaction, including such as interaction of salt bridge and water bridge and directly associate.The anti-C1s antibody of humanization and complement of the disclosure Epitope specificity in C1s albumen combines." specific binding " refers to at least about 10^-7M or higher, such as 5x 10^-7M、10^- ⁸M、5x10^-8M and higher affinity combine." non-specific binding " refers to be below about 10^-7The affinity of M combines, such as with 10^-6M、10^-5M、10^-4The affinity of M etc. combines.

As used herein, term " CDR " or " complementary determining region " be intended to mean in both heavy chain and light chain polypeptide can Become the discontinuous antigen binding site found in area.Kabat et al., CDR are in Kabat et al., J.Biol.Chem.252： 6609-6616(1977)；Kabat et al., U.S.Dept.of Health and Human Services, " Sequences of Proteins of immunological interest " (1991) (herein also referred to as Kabat 1991)；Chothia et al., J.Mol.Biol.196：901-917 (1987) (herein also referred to as Chothia 1987)；And MacCallum et al., J.Mol.Biol.262：Described in 732-745 (1996), wherein this definition includes the overlapping of amino acid residue when being compared to each other Or subset.It defines and uses herein however, being intended to using the CDR that any definition refers to antibody or grafted antibody or its variant Within the scope of term.It is listed in following table 1 defined in each bibliography as referenced above, covers the amino acid of CDR Residue.The CDR described in Fig. 1-8 is defined according to Kabat 1991.

Table 1：CDR is defined

	Kabat¹	Chothia²	MacCallum³
				V_HCDR-1	31-35	26-32	30-35
V_HCDR-2	50-65	53-55	47-58
				V_HCDR-3	95-102	96-101	93-101
V_LCDR-1	24-34	26-32	30-36
				V_LCDR-2	50-56	50-52	46-55
V_LCDR-3	89-97	91-96	89-96

¹Residue numbering according to Kabat above et al. nomenclature, as above

²Residue numbering according to Chothia above et al. nomenclature, as above

³3 residue numberings according to MacCallum above et al. nomenclature, as above

As used herein, term " CDR-L1 ", " CDR-L2 " and " CDR-L3 " refers respectively in light chain variable region One, second and the 3rd CDR.As used herein, term " CDR-H1 ", " CDR-H2 " and " CDR-H3 " refers respectively to weight chain variable The first, second, and third CDR in area.As used herein, term " CDR-1 ", " CDR-2 " and " CDR-3 " refers respectively to appoint First, second, and third CDR of the variable region of one chain.

As used herein, term " frame " (" FR ") refer to antibody variable region using when be intended to mean in antibody variable All amino acid residues in area outside CDR region.Variable framework is typically length between about 100-120 amino acid Discontinuous amino acid sequence, but be intended only to refer to the amino acid those of outside CDR.As used herein, term " framework region " It is intended to mean each structural domain that frame is separated by CDR.Light chain variable region (areas VL) can have there are four framework region：FR1、FR2、 FR3 and FR4.Similarly, heavy chain variable region (VH) can have there are four framework region：FR1, FR2, FR3 and FR4.

" separation " antibody is identified and the component from its natural environment separates and/or the antibody of recycling.It is certainly The pollution components of right environment are the diagnosis that can interfere antibody or the substance of therapeutical uses, and may include enzyme, hormone and other eggs White matter or nonproteinaceous solute.In some embodiments, antibody will be purified (1) as being measured by Lowry methods, press Antibody weight meter reaches higher than 90%, higher than 95% or higher than 98%, such as by weight of from more than 99%, and (2), which reach, to be enough to lead to Cross the degree at least 15 residues that N-terminal or internal amino acid sequence are obtained using spinning cup sequenator；Or (3) pass through dodecane Base sodium sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) uses Coomassie blue under reduction or non reducing conditions (Coomassie blue) or Silver stain, reach homogeneous.The antibody of separation includes the antibody iM situ in recombinant cell, because of antibody At least one component of natural environment will be not present.In some cases, pass through the anti-of at least one purification step preparative separation Body.

Term " polypeptide ", " peptide " and " protein ", is used interchangeably herein, and refers to the poly- of the amino acid of any length Conjunction form, the amino acid that may include genetic coding and non-genetic coding, the ammonia through chemistry or biochemical modification or derivatization Base acid, and with the polypeptide through modifying peptide backbone.The term includes fusion protein, including but not limited to has allogeneic amino acid sequence The fusion protein of row；With heterologous and homologous leader sequences, with or without the fusion protein of N-terminal methionine residues；It is immune labeled Albumen；Etc..

As used herein, term " treatment " etc. refers to obtaining required pharmacology and/or physiological effect.The effect is complete Or the aspect of partial prophylaxis disease or its symptom can be preventative and/or partially or completely cure disease and/or can return Because that can be therapeutic in terms of the adverse effect of the disease." treatment " covering is to mammal as used in this article, especially It is any treatment of the disease in people, and includes：(a) prevent disease can be easy to illness but be not yet diagnosed as illness by Occur in examination person；(b) inhibit disease, that is, prevent its development；And (c) alleviate disease, that is, cause disease regression.

Term " individual ", " subject ", " host " and " patient ", is used interchangeably herein, refers to mammal, packet It includes but is not limited to muroid (rat, mouse), non-human primates, people, dog, cat, ungulate (for example, horse, ox, sheep, pig, mountain Sheep etc.).These terms are also contemplated by any animal with complement system, such as mammal, fish and some invertebrates.Together These terms of sample include mammal containing complement system, fish and without vertebra companion animals, agricultural animal, labour with animal, dynamic Object garden animal and laboratory animal.

" therapeutically effective amount " or " effective quantity " refers to that anticomplement C1s antibody is being administered to mammal or other subjects use When treating disease, it is sufficient to realize to the amount of the such treatment of the disease." therapeutically effective amount " will according to anticomplement C1s antibody, The age of disease and its severity and subject to be treated, weight etc. and change.

" biological sample " is covered the various samples type obtained from individual and be can be used in diagnosis or detection assay.This is fixed Justice cover biological source blood and other fluid samples, solid tissue sample such as biopsy sample or tissue culture or be originated from its In cell and its offspring.This definition further include after its acquirement, in any way for example by being handled with reagent, it is molten The sample for solving or being enriched with certain components (such as polynucleotides) to operate.Term " biological sample " covers clinical sample, and also Including cell, cell supernatant, cell lysates, serum, blood plasma, biofluid and the tissue sample in culture.Term " biological sample " includes urine, saliva, celiolymph, interstitial fluid, intraocular liquid, synovia, blood constituent such as blood plasma and serum, etc. Deng.Term " biological sample " further includes solid tissue sample, tissue culture sample and cell sample.

Before further describing the invention, it should be understood that the present invention is not limited to the particular embodiments of description, thus certainly It can change.It will also be understood that purpose of the terms used herein just for the sake of description particular embodiment, rather than it is intended that limitation Property, because the scope of the present invention is limited only by the appended claims.

When the range of offer value, it should be understood that unless the context clearly indicates otherwise, otherwise between the range bound Each intervening value, to lower limit unit 1/10th and prescribed limit in any other specified value or intervening value, cover In the present invention.These small range of bounds can be independently include in smaller range, and be also covered by within the present invention, with Any limit value clearly excluded is condition in prescribed limit.When prescribed limit includes one or two limit value, those are eliminated The range of either one or two of included limit value is also included in the present invention.

Unless otherwise defined, all technical and scientific terms used herein all have it is general in fields of the present invention The identical meanings that logical technical staff is generally understood.Although the present invention practice or experiment in can also be used with it is described herein that A little similar or equivalent any methods and material, but what be will now be described is preferred method and material.All publication being mentioned herein Object is hereby incorporated herein by the method quoted together with publication with disclosure and description and/or material.

It must be noted that as used herein and in the appended claims, it is unless the context clearly indicates otherwise, otherwise single Number form formula "one", "an" and " (being somebody's turn to do) " include plural referents.Thus, for example, mentioning a kind of " anti-C1s of humanization Antibody " includes a variety of such antibody and mentions " framework region " including mentioning one or more framework regions and art technology Its equivalent, etc. known to personnel.It should be noted also that claim can be drafted to exclude any optional element.Thus, it should Statement is intended as the proprietary terms such as " only ", " only " being used together with to the narration of claim elements, or uses The antecedent basis of " negative " limitation.

It should be appreciated that certain features of for the sake of clarity present invention described in the context of independent embodiment It can be provided in combination in single embodiment.On the contrary, the sheet described in the context of single embodiment for brevity The various features of invention can also provide individually or in any suitable sub-portfolio.Embodiment related to the present invention owns Combination is particularly included by the present invention and disclosed herein, only as individually and specifically disclosed each group of unification Sample.In addition, all sub-portfolios of each embodiment and its element are also particularly included by the present invention and public affairs herein It opens, only as herein individually and specifically disclosing each sub-portfolio.

The publication being discussed herein is provided to disclose before the date of application of the present invention just for the sake of it.Must not will herein Any content be construed as an admission that, due to prior inventions, the present invention haves no right prior to such publication.Further, the publication provided The date of object may differ from the practical publication date, and the practical publication date may need individually to confirm.

It is described in detail

Present disclose provides a kind of humanized antibodies of conjugated complement C1s albumen (that is, humanization anticomplement C1s antibody, sheet Text also referred to as " the anti-C1s antibody of humanization ", " humanization C1s antibody " and " theme antibody ") and include the core for encoding such antibody The nucleic acid of nucleotide sequence.The disclosure additionally provides a kind of composition of the anti-C1s antibody of the humanization comprising the disclosure.The disclosure carries For the method for generating and using the antibody of the disclosure, nucleic acid and composition.Present disclose provides the diseases for the treatment of complement-mediated Or the method for illness, this method are related to the anti-C1s antibody of humanization using the disclosure.

Anticomplement C1s antibody

Present disclose provides humanization anticomplement C1s antibody and include the pharmaceutical composition of such antibody.Complement C1s is that have The target of attraction, because it is in the upstream of complement cascade and with the substrate specificity of narrow range.In some cases Interested is the antibody for specifically binding C1s activated forms, for example, wherein the antibody does not combine inactive form substantially C1s。

The anti-C1s antibody of the disclosure is humanization, for example, one or more of heavy chain variable region and/or light chain variable region A framework region includes the sequence from human immunoglobulin(HIg) frame.

In some cases, the anti-C1s antibody of the disclosure inhibits the complement component C4 cracking that C1s is mediated, for example, passing through suppression The enzymatic activity of the serine protease domain of C1s processed.In some cases, the anti-C1s antibody of the disclosure inhibits C1s to mediate Complement component C2 cracking.In some cases, the anti-C1s antibody of the disclosure inhibits C4 and the C2 cracking of C1s mediations.

The humanization of framework region reduces the risk that antibody causes human anti-mouse antibody (HAMA) response in people.Can carry out The method of art-recognized measurement immune response is to monitor the HAMA responses in particular patient or during clinical test.In the treatment Immunogenic evaluation can be given when method application starts or during entire to the patient that applied humanized antibody.HAMA is measured to answer It answers, for example, being detected in the blood serum sample from patient to humanization therapeutic reagent by using method known to those skilled in the art Antibody, the method includes surface plasma resonance technology (BIACORE) and/or solid-phase enzyme-linked immune determining adsorptions (ELISA) it analyzes.In many cases, the anti-C1s antibody of theme humanization substantially will not cause HAMA to answer in people experimenter It answers.

On CDR conformations and/or the possibility of antigen is combined to influence based on it, selected from the certain of people variable framework residues Amino acid is replaced.The non-natural juxtaposition that mouse CDR region can be changed framework region with people can generate non-native conformation limitation, this is except non-through It crosses and certain amino acid residues is replaced to correct, otherwise binding affinity is caused to be lost.

Selection for substituted amino acid residue can be determined partially by computer modeling.In this field Know certain hardware and softwares of the 3-D view for generating immunoglobulin molecules.Generally, for immunoglobulin chain Or molecular model is generated for its structural domain since dissolving structure.Compare chain to be modeled and dissolves the chain or knot of three-dimensional structure The amino acid sequence similarity in structure domain, and select to show maximal sequence similitude chain or structural domain as structure molecule mould The starting point of type.The selection shared at least chain of 50% sequence identity or structural domain are for modeling, for example, selection is shared at least 60%, the chain or structural domain of at least 70%, at least 80%, at least 90% sequence identity or higher sequence identity are for building Mould.Modify the real amino acid and initial structure in immunoglobulin chain or structural domain of the initial structure of dissolving to allow modeling In those of between have differences.Then the structure of modification is assembled into complicated immunoglobulin.Finally, pass through energy minimum Change and by verifying all atoms each other within suitable distance and bond distance and bond angle refine in the acceptable limit of chemistry Model.

CDR and framework region such as Kabat, Sequences of Proteins of Immunological Interest It (National Institutes of Health, Bethesda, Md., 1987 and 1991) is defined.Chothia et al., J.Mol.Biol.196：901(1987)；Nature 342：878(1989)；And J.Mol.Biol.186：(it is referred to as 651 (1989) For " Chothia ") propose alternate configurations definition.When the Framework residues as defined in Kabat above are constituted as above Defined in Chothia when structure ring residue, it is humanized antibody that amino acid substitution present in mouse antibodies, which may be selected,." with CDR region is adjacent " residue be included in close to the position of one or more of the primary sequence of Humanized immunoglobulin chain CDR On, for example, in the amino acid residue (ginseng close to the CDR as defined in Kabat or on the position of the CDR as defined in Chothia See for example, Chothia and Lesk JMB 196：901(1987)).These amino acid are especially possible to and the amino acid phase in CDR Interaction also, if be selected from receptor, so that donor CDR is deformed and reduce affinity.Moreover, adjacent amino acid can directly with Antigen interactions (Amit et al., Science, 233：747 (1986)) and select these amino acid from donor can be with It is desirably maintained in all antigen contacts that affinity is provided in original antibodies.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes at least one humanization V_HFramework region.At some In the case of, the anti-C1s antibody of the disclosure includes at least one humanization V_LFramework region.In some cases, the anti-C1s of the disclosure Antibody includes at least one humanization V_HFramework region and at least one humanization V_LFramework region.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes VL CDR present in following amino acid sequence：

DIVLTQSPASLAVSLGQRATISCKASQSVDYDGDSYMNWYQQKTGQPPKILIYDASNLESGIPARFSGSGSGTDFTL NIHPVEEEDAAIYYCQQSNEDPWTFGGGTKLEIK(SEQ ID NO：7).In some cases, the humanization of the disclosure is anti- C1s antibody includes VH CDR present in following amino acid sequence：

EVQLQQSGAELVRPGASVKLSCTASGFNIKDDYIHWVKQRPEQGLEWIGRIDPADGHTKYAPKFQVKATITADTSSN TAYLQLSSLTSEDTAVYYCARYGYGREVFDYWGQGTTLTVSS(SEQ ID NO：8).In some cases, the disclosure The anti-C1s antibody of humanization includes SEQ ID NO：VL CDR and SEQ ID NO present in 7：VH CDR present in 8.

VL CDR1(CDR-L1)：SEQ ID NO：1：KASQSVDYDGDSYMN

VL CDR2(CDR-L2)：SEQ ID NO：2：DASNLES

VL CDR3(CDR-L3)：SEQ ID NO：3：QQSNEDPWT

VH CDR1(CDR-H1)：SEQ ID NO：4：DDYIH

VH CDR2(CDR-H2)：SEQ ID NO：5：RIDPADGHTKYAPKFQV

VH CDR3(CDR-H3)：SEQ ID NO：6：YGYGREVFDY

In some cases, the anti-C1s antibody of the humanization of the disclosure includes to contain cdr amino acid sequence SEQ ID NO：1、 SEQ ID NO：2 and SEQ ID NO：The light chain variable region of 3 (being respectively CDR-L1, CDR-L2 and CDR-L3).

In some embodiments, the anti-C1s antibody of the disclosure includes to contain cdr amino acid sequence SEQ ID NO：4、 SEQ ID NO：5 and SEQ ID NO：The heavy chain variable region of 6 (being respectively CDR-H1, CDR-H2 and CDR-H3).

In some cases, the anti-C1s antibody of the humanization of the disclosure includes to contain the following sequence of areas VH：

(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T) SEDTAVYYCARYGYGREVFDYWGQGTTVTVSS(SEQ ID NO：26).

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VH, it includes in Fig. 1 describe and SEQ ID NO：The amino acid sequence listed in 10 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 1 are Glu, and amino acid 5 is Val, and amino acid 11 is Leu, amino acid 12 For Lys, amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, and amino acid 38 is Lys, and amino acid 40 is Ala, Amino acid 42 is Gly, and amino acid 67 is Ala, and amino acid 75 is Thr, and amino acid 76 is Asn, and amino acid 80 is Leu, amino acid 81 be Gln, and amino acid 83 is Thr and amino acid 1 09 is Val, and wherein the number of amino acid is as depicted in fig. 1.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VH, and it includes describe and SEQ ID in Fig. 1 NO：The amino acid sequence listed in 10.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VH, it includes in Fig. 2 describe and SEQ ID NO：The amino acid sequence listed in 12 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 1 are Glu, and amino acid 5 is Val, and amino acid 11 is Val, amino acid 12 For Lys, amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, and amino acid 38 is Lys, and amino acid 40 is Ala, Amino acid 42 is Gly, and amino acid 67 is Ala, and amino acid 75 is Thr, and amino acid 76 is Asn, and amino acid 80 is Leu, amino acid 81 be Glu, and amino acid 83 is Arg and amino acid 1 09 is Val, and the number of wherein amino acid is as depicted in Figure 2.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VH, and it includes describe and SEQ ID in Fig. 2 NO：The amino acid sequence listed in 12.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VH, it includes in Fig. 3 describe and SEQ ID NO：The amino acid sequence listed in 14 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 1 are Gln, and amino acid 5 is Val, and amino acid 11 is Val, amino acid 12 For Lys, amino acid 13 is Lys, and amino acid 20 is Leu, and amino acid 23 is Thr, and amino acid 38 is Lys, and amino acid 40 is Ala, Amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, and amino acid 76 is Ser, and amino acid 80 is Leu, amino acid 81 be Glu, and amino acid 83 is Arg and amino acid 1 09 is Val, and wherein the number of amino acid is as depicted in fig. 3.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VH, and it includes describe and SEQ ID in Fig. 3 NO：The amino acid sequence listed in 14.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VH, it includes in Fig. 4 describe and SEQ ID NO：The amino acid sequence listed in 16 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 1 are Gln, and amino acid 5 is Val, and amino acid 11 is Val, amino acid 12 For Lys, amino acid 13 is Lys, and amino acid 20 is Val, and amino acid 23 is Thr, and amino acid 38 is Arg, and amino acid 40 is Ala, Amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, and amino acid 76 is Ser, and amino acid 80 is Met, amino acid 81 be Glu, and amino acid 83 is Arg and amino acid 1 09 is Val, and wherein the number of amino acid is as depicted in fig. 4.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VH, and it includes describe and SEQ ID in Fig. 4 NO：The amino acid sequence listed in 16.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VH, it includes in Fig. 5 describe and SEQ ID NO：The amino acid sequence listed in 18 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 1 are Gln, and amino acid 5 is Val, and amino acid 11 is Val, amino acid 12 For Lys, amino acid 13 is Lys, and amino acid 20 is Val, and amino acid 23 is Ala, and amino acid 38 is Arg, and amino acid 40 is Ala, Amino acid 42 is Gly, and amino acid 67 is Val, and amino acid 75 is Thr, and amino acid 76 is Ser, and amino acid 80 is Met, amino acid 81 be Glu, and amino acid 83 is Arg and amino acid 1 09 is Val, and wherein the number of amino acid is as depicted in figures 5.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VH, and it includes describe and SEQ ID in Fig. 5 NO：The amino acid sequence listed in 18.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes to contain the following sequence of areas VL：

DIVLTQSPDSLAVSLGERATISCKASQSVDYDGDSYMNWYQQK(T/P)GQPPK(I/L) LIYDASNLESGIPARFSGSGSGTDFTLTISSLE(E/P)EDFA(I/V)YYCQQSNEDPWTFGGGTKVEIK(SEQ ID NO：27).

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VL, it includes in Fig. 6 describe and SEQ ID NO：The amino acid sequence listed in 20 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 9 are Asp, and amino acid 17 is Glu, and amino acid 40 is Thr, amino acid 46 be Ile, and amino acid 74 is Thr, and amino acid 76 is Ser, and amino acid 77 is Ser, and amino acid 78 is Leu, and amino acid 80 is Glu, amino acid 83 are Phe, and amino acid 85 is Ile and amino acid 1 04 is Val, and wherein the number of amino acid is as depicted in figure 6.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VL, and it includes describe and SEQ ID in Fig. 6 NO：The amino acid sequence listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VL, it includes in Fig. 7 describe and SEQ ID NO：The amino acid sequence listed in 22 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 9 are Asp, and amino acid 17 is Glu, and amino acid 40 is Pro, amino acid 46 be Ile, and amino acid 74 is Thr, and amino acid 76 is Ser, and amino acid 77 is Ser, and amino acid 78 is Leu, and amino acid 80 is Pro, amino acid 83 are Phe, and amino acid 85 is Ile and amino acid 1 04 is Val, and wherein the number of amino acid is as depicted in figure 7.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VL, and it includes describe and SEQ ID in Fig. 7 NO：The amino acid sequence listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure include the areas VL, it includes in Fig. 8 describe and SEQ ID NO：The amino acid sequence listed in 24 has at least 90%, at least 95%, at least 98%, at least 99% or 100% amino The amino acid sequence of acid sequence identity, wherein amino acid 9 are Asp, and amino acid 17 is Glu, and amino acid 40 is Pro, amino acid 46 be Leu, and amino acid 74 is Thr, and amino acid 76 is Ser, and amino acid 77 is Ser, and amino acid 78 is Leu, and amino acid 80 is Pro, amino acid 83 are Phe, and amino acid 85 is Val and amino acid 1 04 is Val, and wherein the number of amino acid is as depicted in fig. 8.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes the areas VL, and it includes describe and SEQ ID in Fig. 8 NO：The amino acid sequence listed in 24.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 1：In 10 1 amino acid sequence of VH variants listed；And b) in Fig. 6 describe and SEQ ID NO：1 amino acid sequence of VL variants listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 1：In 10 1 amino acid sequence of VH variants listed；And b) in Fig. 7 describe and SEQ ID NO：2 amino acid sequence of VL variants listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 1：In 10 1 amino acid sequence of VH variants listed；And b) in Fig. 8 describe and SEQ ID NO：5 amino acid sequence of VL variants listed in 24.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 2：In 12 2 amino acid sequence of VH variants listed；And b) in Fig. 6 describe and SEQ ID NO：1 amino acid sequence of VL variants listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 2：In 12 2 amino acid sequence of VH variants listed；And b) in Fig. 7 describe and SEQ ID NO：2 amino acid sequence of VL variants listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 2：In 12 2 amino acid sequence of VH variants listed；And b) in Fig. 8 describe and SEQ ID NO：5 amino acid sequence of VL variants listed in 24.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 3：In 14 3 amino acid sequence of VH variants listed；And b) in Fig. 6 describe and SEQ ID NO：1 amino acid sequence of VL variants listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 3：In 14 3 amino acid sequence of VH variants listed；And b) in Fig. 7 describe and SEQ ID NO：2 amino acid sequence of VL variants listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 3：In 14 3 amino acid sequence of VH variants listed；And b) in Fig. 8 describe and SEQ ID NO：5 amino acid sequence of VL variants listed in 24.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 4：In 16 4 amino acid sequence of VH variants listed；And b) in Fig. 6 describe and SEQ ID NO：1 amino acid sequence of VL variants listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 4：In 16 4 amino acid sequence of VH variants listed；And b) in Fig. 7 describe and SEQ ID NO：2 amino acid sequence of VL variants listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 4：In 16 4 amino acid sequence of VH variants listed；And b) in Fig. 8 describe and SEQ ID NO：5 amino acid sequence of VL variants listed in 24.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 5：In 18 5 amino acid sequence of VH variants listed；And b) in Fig. 6 describe and SEQ ID NO：1 amino acid sequence of VL variants listed in 20.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 5：In 18 5 amino acid sequence of VH variants listed；And b) in Fig. 7 describe and SEQ ID NO：2 amino acid sequence of VL variants listed in 22.

In some cases, the anti-C1s antibody of the humanization of the disclosure includes：A) describe and SEQ ID NO in Fig. 5：In 18 5 amino acid sequence of VH variants listed；And b) in Fig. 8 describe and SEQ ID NO：5 amino acid sequence of VL variants listed in 24.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure is combined from the individual with complement system Complement C1s albumen.In some embodiments, the anti-C1s antibody of the humanization of the disclosure is combined from the food in one's mouth with complement system The complement C1s albumen of newborn animal, fish or invertebrate.In some embodiments, the anti-C1s antibody of the humanization of the disclosure In conjunction with mammal complement C1s albumen.In some embodiments, the anti-C1s antibody combination people complement C1s of the humanization of the disclosure Albumen.In some embodiments, the anti-C1s antibody combination rat complement C1s albumen of the humanization of the disclosure.In some embodiment party In case, the anti-C1s antibody of humanization of the disclosure is combined with amino acid sequence (the SEQ ID NO described in Figure 13：9) complement C1s albumen.Amino acid sequence SEQ ID NO：9 indicate homo sapiens (Homo sapiens) complement C1s albumen, listed by Figure 13 Amino acid sequence.

In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, dissociation constant (K_D) No more than 2.5nM.In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_DIt is no more than 2nM.In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_DNo more than 1nM.One In a little embodiments, the anti-C1s antibody conjugated complement C1s albumen of humanization of the disclosure, K_DNo more than 0.9nM, it is no more than 0.8nM, no more than 0.7nM, no more than 0.6nM, no more than 0.5nM, no more than 0.4nM, no more than 0.3nM, be no more than 0.2nM, it is no more than 0.1nM.In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_D No more than 0.3nM.In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_DIt is no more than 0.2nM.In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_DNo more than 0.1nM. The method that the combination for measuring antibody and C1s albumen can be determined by those skilled in the art.

In some embodiments, the anti-C1s antibody conjugated complement C1s albumen of the humanization of the disclosure, K_DNo more than 90pM, No more than 80pM, be no more than 70pM, be no more than 60pM, be no more than 50pM, be no more than 40pM, be no more than 30pM, be no more than 20pM, No more than 10pM, no more than 9pM, no more than 8pM, no more than 7pM, no more than 6pM, no more than 5pM, no more than 4pM, be no more than 3pM, it is no more than 2pM, is no more than 1pM.

In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, dissociation constant (K_D) it is no more than 2.5nM.In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_DNo More than 2nM.In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_DIt is no more than 1nM.In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_DNo more than 0.9nM, No more than 0.8nM, no more than 0.7nM, no more than 0.6nM, no more than 0.5nM, no more than 0.4nM, no more than 0.3nM, do not surpass It crosses 0.2nM, be no more than 0.1nM.In some embodiments, the anti-C1s antibody combination people complement C1s eggs of the humanization of the disclosure In vain, K_DNo more than 0.3nM.In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_D No more than 0.2nM.In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_DDo not surpass Cross 0.1nM.The method that the combination for measuring antibody and people's C1s albumen can be determined by those skilled in the art.In some embodiment party In case, the K between antibody and people's C1s albumen is measured using the binding assay as described in embodiment_D。

In some embodiments, the anti-C1s antibody combination people complement C1s albumen of the humanization of the disclosure, K_DIt is no more than 90pM, no more than 80pM, no more than 70pM, no more than 60pM, no more than 50pM, no more than 40pM, no more than 30pM, be no more than 20pM, be no more than 10pM, be no more than 9pM, be no more than 8pM, be no more than 7pM, be no more than 6pM, be no more than 5pM, be no more than 4pM, No more than 3pM, it is no more than 2pM, no more than 1pM.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure inhibits classic complement approach, half maximum suppression Concentration (IC₅₀) it is 10^-8M or lower, 5x10^-9M or lower or 10^-9M or lower.

Nucleic acid, expression vector and host cell

Present disclose provides the nucleic acid of the nucleotide sequence comprising the anti-C1s antibody of humanization disclosed in code book.At some In the case of, the nucleic acid of the disclosure includes the nucleotide sequence in the areas VH of the anti-C1s antibody of humanization disclosed in code book.In some feelings Under condition, the nucleic acid of the disclosure includes the nucleotide sequence in the areas VL of the anti-C1s antibody of humanization disclosed in code book.In some cases Under, the nucleic acid of the disclosure includes the nucleotide sequence in the areas VH and the areas VL of the anti-C1s antibody of humanization disclosed in code book.

The nucleotide sequence of the anti-C1s antibody of humanization disclosed in code book can such as start with one or more controlling elements Son and enhancer be operably connected, controlling element allow the nucleotides sequence be listed in expected target cell (for example, it is genetically modified with Synthesize the cell of encoded antibody) in expression.Therefore, in some cases, present disclose provides comprising disclosed in code book The nucleic acid of the nucleotide sequence of the anti-C1s antibody of humanization, the wherein nucleotide sequence and one or more controlling elements, such as start Son and/or enhancer are operably connected.

Suitable promoter and enhancer element are known in the art.For the suitable promoter in prokaryotic host cell Including but not limited to phage t7 RNA polymerase promoter；T3 promoters；T5 promoters；λ P promoters；Trp promoters；lac Operon promoter；Hybrid promoter, such as lac/tac hybrid promoters, tac/trc hybrid promoters, trp/lac start Son, T7/lac promoters；Trc promoters；Tac promoters etc.；Gpt promoters；AraBAD promoters；Regulation and control promoter in vivo, Such as ssaG promoters or promoter related (see, for example, U.S. Patent Publication the 20040131637th), pagC promoters (Pulkkinen and Miller, J.Bacteriol., 1991：173(1)：86-93；Alpuche-Aranda et al., PNAS, 1992；89(21)：10079-83), nirB promoters (Harborne et al. (1992) Mol.Micro.6：2805-2813) etc. (see, for example, Dunstan et al. (1999) Infect.Immun.67：5133-5141；McKelvie et al. (2004) Vaccine 22：3243-3255；And Chatfield et al., (1992) Biotechnol.10：888-892)；70 promoters of σ, Such as shared 70 promoters of σ (see, for example, GenBank Accession AX798980, AX798961 and AX798183)；Solid phase starts Son, such as dps promoters, spv promoters etc.；Promoter from pathogenicity island SPI-2 (see, for example, WO96/17951)； ActA promoters are (see, for example, Shetron-Rama et al. (2002) Infect.Immun.70：1087-1096)；RpsM starts Son is (see, for example, Valdivia and Falkow (1996) .Mol.Microbiol.22：367)；Tet promoters (see, for example, Hillen, W. and Wissmann, A. (1989) is in Saenger, W. and Heinemann, U. (editor), Topics in In Molecular and Structural Biology, Protein-Nucleic Acid Interaction.Macmillan, London, UK, volume 10,143-162 pages of ground)；SP6 promoters are (see, for example, Melton Et al. (1984) Nucl.Acids Res.12：7035) etc..For prokaryotes such as Escherichia coli (Escherichia coli) In suitable strong promoter include but not limited to Trc, Tac, T5, T7 and P_λ.For the operator in bacterial host cell (operator) non-limiting examples include that (LacI aporepressors change structure to Lac operon operator when being contacted with lactose As to prevent LacI aporepressors binding operation), (with tryptophan compound tense, TrpR is checked trp promoter operator Albumen has the conformation of binding operation；There is no tryptophan, TrpR aporepressors have not binding operation Conformation) and tac promoters operator (see, for example, deBoer et al. (1983) Proc.Natl.Acad.Sci.U.S.A.80：21-25).

In some embodiments, for example, in order to be expressed in yeast cells, suitable promoter is constitutive promoter Such as ADH1 promoters, PGK1 promoters, ENO promoters, PYK1 promoters；Or controllable promoter such as GAL1 promoters, GAL10 promoters, ADH2 promoters, PHO5 promoters, CUP1 promoters, GAL7 promoters, MET25 promoters, MET3 start Son, CYC1 promoters, HIS3 promoters, ADH1 promoters, PGK promoters, GAPDH promoters, ADC1 promoters, TRP1 start Son, URA3 promoters, LEU2 promoters, ENO promoters, TP1 promoters and AOX1 are (for example, be used for pichia (Pichia) in).

In order to be expressed in eukaryocyte, suitable promoter includes but not limited to light chain and/or heavy chain immunoglobulin Gene promoter and enhancer element；Cytomegalovirus pole early promoter；Herpes simplex virus thymidine kinase promoter；In early days With late period SV40 promoter；The promoter being present in the long terminal repeats from retrovirus；Mouse metallothionein - I promoters in vain；With various tissue-specific promoters known in the art.

The selection of suitable carrier and promoter is completely in ordinary skill horizontal extent.

Including the nucleic acid of the nucleotide sequence of the anti-C1s antibody of humanization disclosed in code book may be present in expression vector and/ Or in cloning vector.Present disclose provides the recombinant vector comprising nucleic acid, the nucleic acid includes that humanization disclosed in code book is anti- The nucleotide sequence of C1s antibody, wherein the recombinant vector is cloning vector.Present disclose provides the recombination loads comprising nucleic acid Body, the nucleic acid includes the nucleotide sequence of the anti-C1s antibody of humanization disclosed in code book, wherein the recombinant vector is expression Carrier, for example, the wherein described nucleotide sequence is operably connected to ensure to be compiled with regulating and controlling sequence appropriate in expression vector The expression of the antibody of code.When theme antibody includes two independent polypeptides, the nucleic acid of two polypeptides of coding can be identical or independent Carrier in clone to form one or more recombinant vectors.Recombinant vector may include selected marker, replication orgin and provide weight The other features of duplication and/or the maintenance of group carrier (for example, recombinant expression carrier).

A large amount of suitable carriers and promoter known to those skilled in the art；It is many commercially available for generating theme Recombinant vector.Following carrier is provided by way of example.Bacterium：pBs、phagescript、PsiX174、pBluescript SK, pBs KS, pNH8a, pNH16a, pNH18a, pNH46a (Stratagene, La Jolla, Calif., USA)；pTrc99A、 PKK223-3, pKK233-3, pDR540 and pRIT5 (Pharmacia, Uppsala, Sweden).Eucaryote：pWLneo、 PSV2cat, pOG44, PXR1, pSG (Stratagene) pSVK3, pBPV, pMSG and pSVL (Pharmacia).

Expression vector usually has the suitable restriction site being located near promoter sequence to provide encoding heterologous albumen The insertion of nucleic acid sequence.The effective selected marker in expressive host may be present.Suitable expression vector includes but not limited to disease Poisonous carrier.The example of viral vectors includes but not limited to the viral vectors based on following virus：Vaccinia virus；Polio disease Poison；Adenovirus is (see, for example, Li et al. people, Invest Opthalmol Vis Sci 35：2543 2549,1994；Borras etc. People, Gene Ther 6：515 524,1999；Li and Davidson, PNAS 92：7700 7704,1995；Sakamoto et al., H Gene Ther 5：1088 1097,1999；WO 94/12649；WO 93/03769；WO 93/19191；WO 94/28938； WO 95/11984 and WO 95/00655)；Adeno-associated virus is (see, for example, Ali et al., Hum Gene Ther 9：81 86, 1998；Flannery et al., PNAS 94：6916 6921,1997；Bennett et al., Invest Opthalmol Vis Sci 38：2857 2863,1997；Jomary et al., Gene Ther 4：683 690,1997；Rolling et al., Hum Gene Ther 10：641 648,1999；Ali et al., Hum Mol Genet 5：591 594,1996；Srivastava is in WO 93/ In 09239, Samulski et al., J.Vir. (1989) 63：3822-3828；Mendelson et al., Virol. (1988) 166： 154-165；And Flotte et al., PNAS (1993) 90：10613-10617)；SV40；Herpes simplex virus；Retrovirus vector Body (for example, murine leukemia virus, spleen necrosis virus, and it is originated from retrovirus such as Rous sarcoma virus (Rous Sarcoma Virus), harvey sarcoma virus Harvey Sarcoma Virus), avian leukosis virus, human immunodeficiency virus is (referring to example Such as, Miyoshi et al., PNAS 94：10319 23,1997；Takahashi et al., J Virol 73：7812 7816, 1999), the carrier of Myeloproliferative Sarcoma virus and tumor) etc..

Host cell

Present disclose provides the genetic modification host cells (for example, cell in vitro) of separation, are repaiied with theme nucleic acid heredity Decorations.In some embodiments, the genetic modification host cell of theme separation can generate theme antibody.Such cell is known as " weight Group cell " or " genetic modification host cell ".The genetic modification host cell of the disclosure includes nucleic acid, and the nucleic acid includes coding The nucleotide sequence of the anti-C1s antibody of humanization of the disclosure.

Suitable host cell includes eukaryotic host cell, such as mammalian cell, insect host cell, yeast cells； And prokaryotic cell, such as bacterial cell.Introducing of the theme nucleic acid to host cell can be with for example, pass through calcium phosphate precipitation, the Portugals DEAE What glycan mediated transfect, liposome-mediated transfection, electroporation or other known method are realized.

Suitable mammalian cell includes primary cell and immortalized cell line.Suitable mammal cell line includes Human cell line, non-human primate cells system, rodent (for example, mouse, rat) cell line etc..Suitable mammalian cell System includes but not limited to HeLa cells (for example, American Type Culture Collection (ATCC) number CCL-2), Chinese hamster ovary celI (example Such as, ATCC numbers CRL9618, CCL61, CRL9096), 293 cells (for example, ATCC number CRL-1573), Vero cells, NIH 3T3 cells (for example, ATCC number CRL-1658), Huh-7 cells, bhk cell (for example, ATCC number CCL10), PC12 cells (ATCC number CRL1721), COS cells, COS-7 cells (ATCC number CRL1651), RAT1 cells, mouse Lcell (ATCC Number CCLI.3), human embryo kidney (HEK) (HEK) cell (ATCC number CRL1573), HLHepG2 cells etc..In some cases, described Cell is HEK cells.In some cases, the cell be Chinese hamster ovary celI, such as CHO-K1 cells (ATCC number CCL-61), CHO-M cells, CHO-DG44 cells (ATCC number PTA-3356) etc..In some embodiments, host cell is that COS is thin Born of the same parents.In some embodiments, host cell is 293 cells.In some embodiments, host cell is Chinese hamster ovary celI.

Suitable yeast cells includes but not limited to that pichia pastoris yeast (Pichia pastoris), Finland finish red ferment Female (Pichia finlandica), happiness trehalose Pichia pastoris (Pichia trehalophila), Pichia koclamae, Pichia membranaefaciens (Pichia membranaefaciens), Pichia opuntiae, Pichia thermotolerans, Willow Pichia pastoris (Pichia salictaria), Pichia guercuum, Pi Jiepu Pichia pastoris (Pichia pijperi), Pichia stipitis (Pichia stiptis), pichia methanolica (Pichia methanolica), pichia It is (Pichia sp.), saccharomyces cerevisiae (Saccharomyces cerevisiae), saccharomyces (Saccharomyces sp.), more Shape Hansenula yeast (Hansenula polymorpha), Kluyveromyces (Kluyveromyces sp.), Kluyveromyces Lactis tie up ferment Female (Kluyveromyces lactis), candida albicans (Candida albicans), aspergillus nidulans (Aspergillus Nidulans), aspergillus niger (Aspergillus niger), aspergillus oryzae (Aspergillus oryzae), trichoderma reesei (Trichoderma reesei), Chrysosporium lucknowense, Fusarium (Fusarium sp.), cereal sickle Spore bacterium (Fusarium gramineum), golden yellow Fusariumsp (Fusarium venenatum), Neurospora crassa (Neurospora crassa), Chlamydomonas reinhardtii (Chlamydomonas reinhardtii) etc..In some embodiments, place Chief cell is saccharomyces.In some embodiments, host cell is pichia.

Suitable prokaryotic cell includes but not limited to Escherichia coli, bacillus (for example, bacillus subtilis (B.subtilis)), any one of the kinds of experiments room bacterial strain of lactobacillus (Lactobacillus) etc..See, e.g., Carrier et al. (1992) J.Immunol.148：1176-1181；U.S. Patent No. 6,447,784；And Sizemore et al. (1995)Science 270：299-302.In general, laboratory strains are non-pathogenic bacterial strains.In some embodiments, place Chief cell is Escherichia coli.In some embodiments, host cell is bacillus subtilis.

Pharmaceutical composition

Include the pharmaceutical composition of the anti-C1s antibody of the humanization comprising the disclosure present disclose provides composition.It is general and Speech, pharmaceutical composition, herein also referred to as preparation, including the anti-C1s antibody of the humanization of a effective amount of disclosure." effective quantity " means Be enough to generate the dosage of required result, required result for example, mitigate with the relevant ill symptoms of the disease or illness of complement-mediated, The disease of complement-mediated or the symptom of illness improve, the disease of complement-mediated or the progression of illness etc..In general, required result At least compared with the control, the symptom of the disease of complement-mediated or illness mitigates.In some embodiments, the people source of the disclosure Change anti-C1s antibody it is formulated and/or modification so that antibody can pass through blood-brain barrier.In some embodiments, the disclosure The anti-C1s antibody of humanization by avoid blood-brain barrier it is this in a manner of deliver.In some embodiments, with being conducive to across blood The medicament of brain barrier prepares the anti-C1s antibody of humanization of the disclosure.In some embodiments, the anti-C1s of the humanization of the disclosure Antibody is merged directly or by connector with the compound across blood-brain barrier is promoted.

Preparation

In subject methods, can use can generate any convenient manner of required therapeutic effect or diagnosis effect to place The anti-C1s antibody of the main humanization using the disclosure.Therefore, medicament can be mixed in a variety of preparations for treating application.It is more special Not, can by with pharmaceutically acceptable carrier appropriate, pharmaceutically acceptable diluent or pharmaceutically acceptable Excipient composition the anti-C1s antibody of the humanization of the disclosure is configured to pharmaceutical composition and can be configured to solid, semisolid, The preparation of liquid or gas form, as tablet, capsule, pulvis, granula, ointment, solution, suppository, injection, inhalant are gentle molten Glue.In some embodiments, pharmaceutical composition includes the anti-C1s antibody of humanization of the disclosure and pharmaceutically acceptable figuration Agent.

In pharmaceutical dosage form, the anti-C1s antibody of humanization of the disclosure can be applied in the form of its pharmaceutically acceptable salt With, or also can be used alone or suitably combine and be applied in combination with other medicines reactive compound.Following methods and figuration Agent is merely illustrative and not in any way limiting.

For oral preparation, the anti-C1s antibody of humanization of the disclosure can be used alone or with other suitable additives groups It closes and uses to prepare tablet, pulvis, granula or capsule, such as combine with conventional additives, such as lactose, mannitol, corn shallow lake Powder or potato starch；It is combined with adhesive, such as avicel cellulose, cellulose derivative, gum arabic, cornstarch or bright Glue；It is combined with disintegrant, such as cornstarch, potato starch or sodium carboxymethylcellulose；With lubricant combination, such as talcum or hard Fatty acid magnesium；And if desired, with diluent, buffer, wetting agent, preservative and flavoring agent combination.

It can be by making antibody dissolving, suspending or being emulsifiable in aqueous or non-aqueous solvent, such as plant or other similar oil, the third two Alcohol, synthctic fat acid glyceride, the organosilane ester (e.g., ethyl oleate) of injectable, higher fatty acids or propylene glycol ester in, will The anti-C1s antibody of humanization of the disclosure is configured to the preparation for injection；And if desired, have conventional additives such as solubilizer, Isotonic agent, suspending agent, emulsifier, stabilizer and preservative.Parenteral medium includes sodium chloride solution, woods grignard glucose (Ringer ' s dextrose), glucose and sodium chloride, Lactated Ringer'S Solution or fixed oil.Intravenous vehicles include Fluid and nutritional supplement, electrolyte replenisher (replenishers such as based on woods grignard glucose).In addition, according to pharmaceutical composition The pharmaceutical composition of the desired use of object, the disclosure may include other reagents, such as dopamine or psychopharmacological agents.

It can be by by the anti-C1s antibody of the humanization of the disclosure with the desired purity and optional physiologically acceptable load Body, other excipient, stabilizer, surfactant, buffer and/or tonicity agent mix to prepare the humanization comprising the disclosure The pharmaceutical composition of anti-C1s antibody.Acceptable carrier, other excipient and/or stabilizer are under the dosage of use and concentration It is nontoxic to receptor, and include buffer such as phosphate, citrate and other organic acids；Antioxidant, including ascorbic acid, Glutathione, cysteine, methionine and citric acid；Preservative (such as ethyl alcohol, benzyl alcohol, phenol, metacresol, the first between chlorine Phenol, methyl p-hydroxybenzoate or propylparaben, benzalkonium chloride or combinations thereof)；Amino acid such as arginine, sweet ammonia Acid, ornithine, lysine, histidine, glutamic acid, aspartic acid, isoleucine, leucine, alanine, phenylalanine, junket ammonia Acid, tryptophan, methionine, serine, proline and combinations thereof；Monosaccharide, disaccharides and other carbohydrate；Low molecular weight (less than about 10 residues) polypeptide；Protein, such as gelatin or seralbumin；Chelating agent such as EDTA；Sugar as trehalose, sucrose, Lactose, glucose, mannose, maltose, galactolipin, fructose, sorbose, gossypose, gucosamine, N- methyl glucoses osamine, Galactosamine and neuraminic acid；And/or nonionic surface active agent such as tween (Tween), Brij Pluronics, Triton-X or polyethylene glycol (PEG).

Pharmaceutical composition can be in liquid form, lyophilized form or the liquid form by lyophilized form rehydration, wherein freeze-drying system Sterile solution rehydration is used in agent before administration.It is the pure water of add-back certain volume to make the standardization program of freeze-dried composition rehydration (volume removed during being generally equal to lyophilized)；However include that the solution of antiseptic can be used for producing the drug for parenteral administration Composition；Referring further to Chen (1992) Drug Dev Ind Pharm 18,1311-54.

Exemplary antibodies concentration range in theme pharmaceutical composition can be about 1mg/mL to about 200mg/mL or about 50mg/ ML is to about 200mg/mL, or about 150mg/mL to about 200mg/mL.

The aqueous formulation of the anti-C1s antibody of humanization of the disclosure can be prepared in pH buffer solutions, such as in the range of about 4.0 To about 7.0, or about 5.0 to about 6.0, or optionally under about 5.5 pH.It is suitble to the example of buffer solution of the pH within the scope of this to include Phosphate, histidine, citrate, succinate, acetate buffer and other organic acid buffer agent.According to such as buffer With the required tonicity of preparation, buffer concentration can be about 1mM to about 100mM or about 5mM to about 50mM.

It may include tonicity agent in antibody preparation to adjust the tonicity of preparation.Exemplary tonicity agent includes sodium chloride, chlorination Potassium, glycerine and any component from amino acids, carbohydrate and combinations thereof.In some embodiments, aqueous formulation is isotonic, though Right hypertonic or hypotonic solution may also be suitable.Term " isotonic " refers to some other solution such as physiology that solution has and compares therewith Salting liquid or the identical tonicity of serum.Tonicity agent can be by about 5mM to the amount of about 350mM, such as makes by the amount of 100mM to 350nM With.

Also surfactant can be added into antibody preparation to reduce the aggregation of the antibody of preparation and/or by particle in preparation Formation be minimized and/or reduce absorption.Exemplary surfactants include that polyoxyethylene sorbitan fatty acid ester (is spat Temperature), polyoxyethylene alkyl ether (Brij), polyoxyethylene alkylphenol ether (Triton-X), Pluronic F68 (Poloxamer, Pluronic) and lauryl sodium sulfate (SDS).Suitable polyoxyethylene sorbitan fatty acid ester Example is polysorbate20 (with trade mark Tween 20^TMSale) and polysorbate80 (with trade mark Tween 80^TMSale). The example of suitable Pluronic F68 is with titleF68 or Poloxamer 188^TMSale Those of.The example of suitable polyoxyethylene alkyl ether is with trade mark Brij^TMThose of sale.Surfactant it is exemplary dense Degree may range from about 0.001% to about 1%w/v.

Also freeze drying protectant can be added with protect unstable active constituent (for example, protein) in freeze-drying process from Instability condition.For example, as it is known that freeze drying protectant include sugared (including dextrose and saccharose)；Polyalcohol (including mannitol, mountain Pears sugar alcohol and glycerine)；With amino acid (including alanine, glycine and glutamic acid).It may include the amount of about 10mM to 500nM Freeze drying protectant.

In some embodiments, subject formulations include the disclosure the anti-C1s antibody of humanization and it is one or more more than The medicament (for example, surfactant, buffer, stabilizer, tonicity agent) of identification and one or more anti-corrosions are not conformed to substantially Agent, such as ethyl alcohol, benzyl alcohol, phenol, metacresol, parachlorometacresol, methyl p-hydroxybenzoate or propylparaben, benzene Prick oronain or combinations thereof.In other embodiments, include preservative in the formulation, for example, concentration range is about 0.001 to about 2% (w/v).

For example, subject formulations can be adapted for the liquid or lyophilized preparation of parenteral administration, and may include：About 1mg/mL To the anti-C1s antibody of humanization of the disclosure of about 200mg/mL；About 0.001% to about 1% at least one surfactant；About The buffer of 1mM to about 100mM；The stabilizer of optionally about 10mM to about 500mM；The tonicity agent of about 5mM to about 305mM； And pH is about 4.0 to about 7.0.

For another example, theme parenteral administration be liquid or lyophilized preparation, it includes：This public affairs of about 1mg/mL to about 200mg/mL The anti-C1s antibody of humanization opened；0.04% polysorbas20 w/v；20mM L-Histidines；With 250mM sucrose；And pH is 5.5.

For another example, theme parenteral administration includes lyophilized preparation, it includes：1) the theme antibody of 15mg/mL；0.04% tween 20w/v；20mM L-Histidines；With 250mM sucrose；And pH is 5.5；Or 2) the theme antibody of 75mg/mL；0.04% tween 20w/v；20mM L-Histidines；With 250mM sucrose；And pH is 5.5；Or 3) the theme antibody of 75mg/mL；0.02% tween 20w/v；20mM L-Histidines；With 250mM sucrose；And pH is 5.5；Or 4) the theme antibody of 75mg/mL；0.04% tween 20w/v；20mM L-Histidines；With 250mM trehaloses；And pH is 5.5；Or 5) the theme antibody of 75mg/mL；0.02% spits Warm 20w/v；20mM L-Histidines；With 250mM trehaloses；And pH is 5.5.

For another example, theme parenteral administration is liquid preparation, it includes：1) the theme antibody of 7.5mg/mL；0.02% tween 20w/v；120mM L-Histidines；With 250 125mM sucrose；And pH is 5.5；Or 2) the theme antibody of 37.5mg/mL； 0.02% polysorbas20 w/v；10mM L-Histidines；With 125mM sucrose；And pH is 5.5；Or 3) theme of 37.5mg/mL resists Body；0.01% polysorbas20 w/v；10mM L-Histidines；With 125mM sucrose；And pH is 5.5；Or 4) the theme of 37.5mg/mL Antibody；0.02% polysorbas20 w/v；10mM L-Histidines；With 125mM trehaloses；And pH is 5.5；Or 5) 37.5mg/mL Theme antibody；0.01% polysorbas20 w/v；10mM L-Histidines；With 125mM trehaloses；And pH is 5.5；Or 6) 5mg/mL Theme antibody；0.02% polysorbas20 w/v；20mM L-Histidines；With 250mM trehaloses；And pH is 5.5；Or 7) 75mg/mL Theme antibody；0.02% polysorbas20 w/v；20mM L-Histidines；With 250mM mannitols；And pH is 5.5；Or 8) The theme antibody of 75mg/mL；0.02% polysorbas20 w/v；20mM L-Histidines；With 140mM sodium chloride；And pH is 5.5；Or 9) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v；20mM L-Histidines；With 250mM trehaloses；And pH is 5.5； Or 10) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v；20mM L-Histidines；With 250mM mannitols；And pH It is 5.5；Or 11) the theme antibody of 150mg/mL；0.02% polysorbas20 w/v；20mM L-Histidines；With 140mM sodium chloride；And And pH is 5.5；Or 12) the theme antibody of 10mg/mL；0.01% polysorbas20 w/v；20mM L-Histidines；With 40mM sodium chloride； And pH is 5.5.

Theme antibody can be used for will be via in the aerosol preparation that sucking is applied.It is acceptable that theme antibody can be formulated into pressurization Propellant in, such as dicholorodifluoromethane, propane, nitrogen.Aerosol preparation such as nose spray preparation includes activating agent and anti-corrosion The purification of aqueous solutions or other solution of agent and isotonic agent.Such preparation is adjusted to the pH compatible with schneiderian membrane and isotonic state.

Unit dosage forms for oral administration can be provided, such as syrup, elixir and suspension, wherein each dosage unit, such as One, a soupspoon or a piece of, the composition containing predetermined amount.Similarly, the unit dosage forms for injecting or intravenously applying It may include theme antibody in the composition, in the solution in sterile water, physiological saline or other pharmaceutically acceptable carriers.

As used herein, term " unit dosage forms " refers to the physics for being suitable as the unit dose of humans and animals subject Discrete unit, the anti-C1s antibody of humanization of the disclosure of each unit containing predetermined amount are pharmaceutically acceptable dilute by being enough to combine Release the amount calculating that agent, carrier or medium generate required effect.The specification of theme antibody may depend on use specific antibodies and In effect to be achieved and host with the relevant drug effect of each antibody.

Other administration mode will be also used together with disclosed method.For example, theme antibody can be formulated in suppository simultaneously And in some cases, it is formulated in aerosol and nasal composition.For suppository, medium composition will include tradition Adhesive and carrier such as polyalkylene glycol or triglycerides.Such suppository can by containing in about 0.5% to about 10% (w/w), The mixture of active constituent in the range of for example, about 1% to about 2% is formed.

Nasal formulations, which are typically included both generate schneiderian membrane, stimulates the medium that will not obviously upset fibre function Object.Diluent such as water, brine or other known substance can be used.Nasal formulations can also contain preservative, such as, but not limited to, neoprene Alcohol and benzalkonium chloride.Surfactant may be present to enhance absorption of the schneiderian membrane to theme antibody.

Theme antibody can be used as ejection preparation application.In general, injectable composition is prepared into liquid solution or suspension； The solid form being suitble in liquid vehicle in solution or suspension can be prepared before injection.Preparation also can through emulsification or Antibody is set to be encapsulated in liposome medium.

Suitable excipient vehicles are, such as water, brine, glucose, glycerine, ethyl alcohol etc. and combinations thereof.If in addition, It needs, which can contain a small amount of auxiliary substance such as wetting agent or emulsifier or pH buffer.Prepare the practical side of such dosage form Method is known, or will be will be apparent to persons skilled in the art.Referring to,Such as, Remington ' s Pharmaceutical Sciences, Mack Publishing Company, Easton, Pennsylvania, the 17th edition, 1985.The composition or preparation to be applied will be enough to reach required state in being controlled subject containing a certain amount of anyway Theme antibody.

Pharmaceutically acceptable excipient is easy to for the public to obtain such as medium, adjuvant, carrier or diluent.And And pharmaceutically acceptable auxiliary substance, such as pH adjusting agent and buffer, tonicity contributor, stabilizer, wetting agent, it is easy to It is for the public to obtain.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure is formulated in controlled release preparation.This field can be used Well known method prepares sustained release preparation.The suitable example of sustained release preparation includes partly oozing for the solid hydrophobic polymers containing antibody Saturating matrix, wherein the matrix is in the form of moulded products, such as film or microcapsules.The example of sustained-release matrix include polyester, It is the copolymer of Pidolidone and Pidolidone ethyl ester, nondegradable ethane-acetic acid ethyenyl ester, hydrogel, polyactide, degradable Lactic acid-ethanol copolymer and poly- D- (-) -3-hydroxybutyrate.Additive appropriate can be used, by controlling water content The possible loss of bioactivity of antibody included in sustained release preparation is prevented with by developing particular polymers base composition Change with possible immunogenicity.

Controlled release within the scope of this disclosure may be considered that refer to any one of many extended release dosage forms.In order to The purpose of the disclosure, following term can be considered as and controlled release basic equivalence：Continuous release, controlled release, sustained release, reservoir, delay It discharges, gradually discharges, releases immediately, release for a long time, procedural release, extended release, release, long-time are discharged, stored up in proportion Warehousing, delay, slow graceful release, interval release, sustained release, time coating, time controlled released, delayed-action, extension effect, separation time Effect, long-acting, long term, repeat function, slowly effect, continuous action and sustained release drugs with function.These terms it is further Discuss can in Lesczek Krowczynski,Extended-Release Dosage Forms, 1987 (CRC Press, Inc. it is found in).

Various controlled-release technologies cover very extensive pharmaceutical dosage form.Controlled-release technology includes but not limited to physical system and change System.

Physical system includes but not limited to the reservoir system with rate-controlling membrane, such as microencapsulation (microencapsulation), macrocyst (macroencapsulation) and membranous system；Reservoir system without rate-controlling membrane, such as Hollow fibre, ultramicropore cellulose triacetate and porous polymer matrix and foam；Total system, including physics be dissolved in it is non-porous, System and object those of in polymerization or elastomeric matrices (for example, not aggressive, aggressivity, environmental factor enter and degradable) Reason is dissolved in the material in non-porous, polymerization or elastomeric matrices (for example, not aggressive, aggressivity, environmental factor enter and degradable) Material；Layer structure includes being chemically similar with outer control layer or different reservoir；With other physical methods, such as osmotic pumps, Or it is adsorbed onto on ion exchange resin.

Chemical system includes but not limited to the chemical erosion (for example, uneven or uniform erosion) of polymer substrate, or poly- The bioerosion (for example, uneven or uniform) of polymer matrix.It can be to other discussion of the system classification for controlled release Agis F.Kydonieus,Controlled Release Technologies：Methods, Theory and Applications, find in 1980 (CRC Press, Inc.).

Controlled-release pharmaceutical formulation there are many exploitations for oral administration.These include but not limited to the stomach of osmotic pressure control Intestine delivery system；The stomach and intestine delivery system of hydrokinetic pressure control；The stomach and intestine delivery system of film infiltration control comprising micropore The stomach and intestine delivery system of film infiltration control；The intestinal-specific controlled release stomach and intestine delivery system of anti-gastric juice；The stomach and intestine of gel diffusion control Delivery system；With the stomach and intestine delivery system of ion exchange control comprising cation and anionic drugs.It is passed about controlled release drug Send system other information can in Yie W.Chien,Novel Drug Delivery Systems, 1992 (Marcel Dekker, Inc.) in find.

Dosage

By attending physician or the medical worker of other qualifications suitable dose can be determined based on various clinical factors.Such as medicine Well known in field, the dosage of any patient both depends on many factors, including the size of patient, body surface area, year Age, the specific compound to be applied, Gender, administration time and approach, general health and the other medicines being administered simultaneously Object.It can be by each dosage between 1ng/kg weight and 20mg/kg weight, such as between 0.1mg/kg weight to 10mg/kg Between weight, such as between 0.5mg/kg weight to the amount between 5mg/kg weight using theme antibody；However, it is contemplated that having arrived low In or higher than the example ranges dosage, be especially considering that above-mentioned factor.It, can if the program is continuous infusion With in the range of 1 μ g per minute are to 10mg/kg weight.

In some embodiments, the dosage of the anti-C1s antibody of the humanization of the disclosure is in 0.001 μ g to the range of 1000 μ g It is interior；However, it is contemplated that having arrived the dosage below or above the example ranges, it is especially considering that above-mentioned factor.At some In embodiment, the dosage range can be that for example, about 0.0001 to 100mg/kg weight, or about 0.01 to 5mg/kg weight (for example, 0.02mg/kg, 0.25mg/kg, 0.5mg/kg, 0.75mg/kg, 1mg/kg, 2mg/kg etc.).Such as dosage can be 1mg/kg weight or 10mg/kg weight or within the scope of 1-10mg/kg, or be at least 1mg/kg.Dosage among range above It is also contemplated that within the scope of the present invention.

In some embodiments, it presses and about 1 μ g/ml to about 1mg/ml is provided, for example, about 1 μ g/ml to about 2.5 μ g/ml, About 2.5 μ g/ml to about 5 μ g/ml, about 5 μ g/ml are to about 7.5 μ g/ml, and about 7.5 μ g/ml to about 10 μ g/ml, about 10 μ g/ml are to about 25 μ g/ml, about 25 μ g/ml are to about 50 μ g/ml, and about 50 μ g/ml to about 100 μ g/ml, about 100 μ g/ml are to about 250 μ g/ml, about 250 μ g/ml to about 500 μ g/ml, about 500 μ g/ml are to about 750 μ g/ml or about 750 μ g/ml to the serum peak of about 1000 μ g/ml The amount of concentration, using the anti-C1s antibody of the humanization of the disclosure.In some embodiments, it presses to provide and is higher than 1mg/ml, for example, about The amount of 1mg/ml to about 2mg/ml, about 2mg/ml to about 5mg/ml or about 5mg/ml to the peak serum concentration of about 10mg/ml, application The anti-C1s antibody of theme.It can be according to humanized antibody arbitrary a period of time of any time top application disclosure.

Technical staff will readily recognize that dosage level and administration time table can according to specific antibodies, severity of symptom and Subject changes the neurological susceptibility of side effect.The preferred dose and administration time table that give compound are easy to by this field Technical staff determine by various modes.

Administration method

Using any methods availalbe and suitable for the approach of drug delivery, including internal and external method and whole body drawn game Portion's administration method applies theme antibody to individual.

Conventional and pharmaceutically acceptable administration method includes intranasal, intramuscular, tracheal strips, intrathecal, encephalic, subcutaneous, skin In interior, local, intravenous, peritonaeum, intra-arterial (for example, passing through arteria carotis), spinal cord or brain delivering, rectum, nose, take orally and Other enterals and parenteral administration approach.If desired, administration method can combine, or adjusted according to antibody and/or required effect. Theme antibody compositions can be in that single dose or multi-dose are applied.In some embodiments, theme antibody compositions are administered orally. In some embodiments, theme antibody compositions are applied via inhalation route.In some embodiments, intranasal administration theme Antibody compositions.In some embodiments, local application theme antibody compositions.In some embodiments, encephalic is applied Theme antibody compositions.In some embodiments, theme antibody compositions are intravenously applied.In some embodiments, it passes through Subcutaneous administration theme antibody compositions.In some embodiments, intramuscular applies theme antibody compositions.In some embodiment party It is intrathecal to apply theme antibody compositions in case.

Any available conventional method and approach suitable for Conventional drug delivery, including whole body or localization approach can be used The antibody of the disclosure is applied to host.In general, the administration method considered of the present invention include but is not limited to enteral, Parenteral or inhalation route.

Parenteral administration approach in addition to sucking and applying include but is not limited to part, percutaneous, subcutaneous, intramuscular, Socket of the eye is interior, intracapsular, intraspinal, breastbone is interior, intrathecal and intravenous route, i.e., except through any administration method other than alimentary canal. Parenteral administration can be carried out to realize the whole body or local delivery of theme antibody.When needing systemic delivery, using being usually directed to medicine Invasive or systemic Absorption the part of object preparation or mucosal administration.

Also theme antibody can be delivered to subject by enteral administration.Enteral administration approach is including but not limited to oral It is delivered with rectum (for example, using suppository).

Treatment means the improvement at least with the relevant symptom of pathological condition for tormenting host, is used in a broad sense wherein improving Refer to and is at least dropped with the pathological condition for the treatment of, such as the amplitude of the disease of complement-mediated or the relevant parameter of illness (such as symptom) It is low.Thus, treatment further includes following situations, and wherein pathological condition or at least correlated symptom is totally constrained, such as anti- Only it occurs, or stops, such as terminates so that host is not subjected to the pathological condition, or is at least not subjected to become the pathology The symptom of situation feature.

In some embodiments, by injecting and/or delivering, for example, position into cerebral artery or directly to brain group Knit the anti-C1s antibody of the interior humanization using the disclosure.Theme humanized antibody can also be applied directly to target area, such as pass through base Because rifle is delivered to target area.

A variety of hosts (wherein term " host " with term " subject ", " individual " and " patient " is interchangeable herein makes With) can be treated according to subject methods.Usual such host is " mammal " or " mammal ", and wherein these terms are extensive Ground is used to describe to belong to the organism of Mammalia, including carnivore mesh (for example, cat), herbivore mesh are (for example, ox, Ma He Sheep), omnivorous animal mesh (for example, dog, goat and pig), Rodentia (for example, mouse, cavy and rat) and Primates (for example, People, chimpanzee and monkey).In some embodiments, host is the individual for having complement system, such as mammal, fish or nothing Vertebrate.In some embodiments, host is mammal containing complement system, fish or without vertebra companion animals, agriculture Industry animal, labour animal, zoo animal and laboratory animal.In some embodiments, host behaves.

Embodiment includes including the composition of container, and it includes theme that the container, which is suitable for accommodating for what is applied to individual, The composition of anti-C1s antibody.For example, theme antibody can be placed in the container suitable for accommodating pharmaceutical composition.Container can be example If bottle (for example, having closing device, such as lid), blister package are (for example, each bubble-cap can be provided to one or more dosage Closing), bottle, the flexible package Mylar or polybag of sealing (for example), ampoule (for the single dose in solution), drop Pipe, syringe, film, pipe etc..In some embodiments, container, such as sterile chamber, including theme pharmaceutical composition. Container is bottle or syringe in some embodiments.Container is bottle in some embodiments.Container in some embodiments For syringe.

Provide the examination of the anti-C1s antibody of humanization (such as in oral or injection dosage) of the disclosure with unit dose Agent box.To describe in treating pathological condition of interest in addition to the container for accommodating unit dose in such kit The packaged information specification of the purposes of the antibody and subsidiary benefit.Preferred compound and unit dose be it is above-described that A bit.

The method for treating the disease or illness of complement-mediated

Present disclose provides the methods of the disease or illness for the treatment of complement-mediated.The method is usually directed to in need The anti-C1s antibody of humanization of a effective amount of disclosure of individual application or the pharmaceutical composition for including such antibody.In some cases Under, the activity using complement C1s in cell, tissue, fluid or the organ for adjusting individual of the anti-C1s antibody of theme, and treat The disease or illness of complement-mediated.Present disclose provides the method for inhibiting the activation of complement component C4 in individual, the method packets It includes and applies the anti-C1s antibody of humanization of a effective amount of disclosure to individual or include the pharmaceutical composition of such antibody.The disclosure The active methods of complement C1s inhibited in individual are provided, the method includes the people to a effective amount of disclosure of individual application The anti-C1s antibody of sourceization or the pharmaceutical composition for including such antibody.Present disclose provides reduce in individual (for example, the stream of individual In body, tissue or organ) complement component pyrolysis product horizontal method, the method includes a effective amount of to individual application The anti-C1s antibody of humanization of the disclosure or the pharmaceutical composition for including such antibody.

In some cases, the method for the individual of disease or illness of the treatment of the disclosure with complement-mediated includes to a Body applies the anti-C1s antibody of humanization of a effective amount of disclosure or a effective amount of pharmaceutical composition, described pharmaceutical composition include： A) the anti-C1s antibody of the humanization of the disclosure；With the pharmaceutically acceptable excipient suitable for being applied to such individual.In some realities It applies in scheme, individual is mammal.In some embodiments, individual is people.Using those skilled in the art can be passed through Known any approach, including those disclosed herein approach.In some embodiments, using being intravenous.In some realities It applies in scheme, using being intrathecal.In some embodiments, using being subcutaneous.In some embodiments, using for muscle It is interior.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The horizontal amount of the complement component pyrolysis product of (for example, in the fluid of individual, tissue or organ) in individual.In some cases Under, the theme medicine group of " effective quantity " of the anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure " effective quantity " for closing object is made in individual (for example, the stream of individual to when individual application in need with one or more dosage In body, tissue or organ) complement component pyrolysis product level with not no C1s Antybody therapies anti-with humanization when, such as with The level of complement component pyrolysis product before the anti-C1s Antybody therapies of humanization in fluid, tissue or organ is at least compared to reduction 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, At least 80%, at least 85%, at least 90%, at least 95% or 100% amount.In some embodiments, individual is dynamic for lactation Object.In some embodiments, individual is people.Using can pass through any approach well known by persons skilled in the art, including this Literary those disclosed approach.In some embodiments, using being intravenous.In some embodiments, administration method is sheath It is interior.In some embodiments, administration method is intravenous.In some embodiments, administration method is subcutaneous.In some realities It applies in scheme, administration method is intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The active amount of the classic complement approach of (for example, in the fluid of individual, tissue or organ) in individual.In some cases, originally The theme pharmaceutical composition of " effective quantity " of the disclosed anti-C1s antibody of humanization or the anti-C1s antibody of humanization comprising the disclosure " effective quantity ", be with one or more dosage to when individual application in need, it is small in humanization anti-C1s antibody application about 48 When it is interior, in about 24 hours, in about 12 hours, in about 8 hours or in about 4 hours, make in individual (for example, the fluid of individual, tissue Or in organ) classic complement approach activity with not no C1s Antybody therapies anti-with humanization when, such as with the anti-C1s of humanization The activity of classic complement approach before Antybody therapy in fluid, tissue or organ compared to reduce at least 10%, at least 20%, extremely Few 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount.In some embodiments, individual is mammal.In some implementations In scheme, individual is people.The application of the anti-C1s antibody of humanization can be wrapped by any approach well known by persons skilled in the art Include those disclosed herein approach.In some embodiments, administration method is intrathecal.In some embodiments, using way Diameter is intravenous.In some embodiments, administration method is subcutaneous.In some embodiments, administration method is intramuscular. Any one of a variety of methods can be used to measure the activity level of classic complement approach.As a non-limiting example, may be used The activity of classic complement approach is in vitro measured, for example, by measuring from blood, serum or the plasma sample that individual obtains Classic complement approach activity level.For example, the classic complement approach in blood, serum or plasma sample can in vitro swash It is living, and the amount of complement component pyrolysis product (such as C5b-9) for activating and generating in this way can be measured.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The active amount of the classic complement approach of (for example, in the fluid of individual, tissue or organ) in individual.In some cases, originally The theme pharmaceutical composition of " effective quantity " of the disclosed anti-C1s antibody of humanization or the anti-C1s antibody of humanization comprising the disclosure " effective quantity ", be with one or more dosage to when individual application in need, it is small in humanization anti-C1s antibody application about 48 When it is interior, in about 24 hours, in about 12 hours, in about 8 hours or in about 4 hours, make in individual (for example, the fluid of individual, tissue Or in organ) classic complement approach activity level with not no C1s Antybody therapies anti-with humanization when, such as resisted with humanization The activity level of classic complement approach before C1s Antybody therapies in fluid, tissue or organ compared to reduce at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% Amount.In some embodiments, individual is mammal.In some embodiments, individual is people.The anti-C1s antibody of humanization Application can pass through any approach well known by persons skilled in the art, including those disclosed herein approach.In some implementations In scheme, administration method is intrathecal.In some embodiments, administration method is intravenous.In some embodiments, it applies Approach is subcutaneous.In some embodiments, administration method is intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The active amount of the classic complement approach of (for example, in the fluid of individual, tissue or organ) in individual.In some cases, originally The theme pharmaceutical composition of " effective quantity " of the disclosed anti-C1s antibody of humanization or the anti-C1s antibody of humanization comprising the disclosure " effective quantity ", be with one or more dosage to it is in need individual application when, keep individual in (for example, individual fluid, In tissue or organ) classic complement approach activity level with not no C1s Antybody therapies anti-with humanization when, such as employment source Change the activity level of the classic complement approach before anti-C1s Antybody therapies in fluid, tissue or organ compared to reduce at least 10%, At least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount, wherein keeping reducing by one section of about 4 hours to about 30 days Time (for example, 4 hours to 8 hours, 8 hours to 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days, 14 days to 21 days or 21 days to 30 days).In some embodiments, individual is mammal.In some embodiments, individual is people.Humanization The application of anti-C1s antibody can pass through any approach well known by persons skilled in the art, including those disclosed herein approach. In some embodiments, administration method is intrathecal.In some embodiments, administration method is intravenous.In some embodiment party In case, administration method is subcutaneous.In some embodiments, administration method is intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The active amount of the classic complement approach of (for example, in the fluid of individual, tissue or organ) in individual.In some cases, originally The theme pharmaceutical composition of " effective quantity " of the disclosed anti-C1s antibody of humanization or the anti-C1s antibody of humanization comprising the disclosure " effective quantity ", be with one or more dosage to it is in need individual application when, keep individual in (for example, individual fluid, In tissue or organ) classic complement approach activity level with not no C1s Antybody therapies anti-with humanization when, such as employment source Change the activity level of the classic complement approach before anti-C1s Antybody therapies in fluid, tissue or organ compared to reduce at least 50%, At least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount, wherein keep reduce about 4 hours to about 21 days a period of time (for example, 4 hours to 8 hours, it is 8 hours to 24 small When, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days or 14 days to 21 days).In some embodiments, individual is mammal. In some embodiments, individual is people.The application of the anti-C1s antibody of humanization can pass through well known by persons skilled in the art What approach, including those disclosed herein approach.In some embodiments, administration method is intrathecal.In some embodiments In, administration method is intravenous.In some embodiments, administration method is subcutaneous.In some embodiments, administration method For intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The horizontal amount of the complement component pyrolysis product of (for example, in the fluid of individual, tissue or organ) in individual.In some cases Under, the theme medicine group of " effective quantity " of the anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure " effective quantity " for closing object is applied about in the anti-C1s antibody of humanization to when individual application in need with one or more dosage In 48 hours, in about 24 hours, in about 12 hours, in about 8 hours or in about 4 hours, make in individual (for example, the fluid of individual, In tissue or organ) complement component pyrolysis product level with not no C1s Antybody therapies anti-with humanization when, such as employment source Change the level of the complement component pyrolysis product before anti-C1s Antybody therapies in fluid, tissue or organ compared to reduce at least 10%, At least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount.In some embodiments, individual is mammal. In some embodiments, individual is people.The application of the anti-C1s antibody of humanization can be by well known by persons skilled in the art any Approach, including those disclosed herein approach.In some embodiments, administration method is intrathecal.In some embodiments, Administration method is intravenous.In some embodiments, administration method is subcutaneous.In some embodiments, administration method is Intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The horizontal amount of the complement component pyrolysis product of (for example, in the fluid of individual, tissue or organ) in individual.In some cases Under, the theme medicine group of " effective quantity " of the anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure " effective quantity " for closing object is applied about in the anti-C1s antibody of humanization to when individual application in need with one or more dosage In 48 hours, in about 24 hours, in about 12 hours, in about 8 hours or in about 4 hours, make in individual (for example, the fluid of individual, In tissue or organ) complement component pyrolysis product level with not no C1s Antybody therapies anti-with humanization when, such as employment source Change the level of the complement component pyrolysis product before anti-C1s Antybody therapies in fluid, tissue or organ compared to reduce at least 50%, At least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount.In some embodiments, individual is mammal.In some embodiments, individual is people.Humanization is anti- The application of C1s antibody can pass through any approach well known by persons skilled in the art, including those disclosed herein approach.One In a little embodiments, administration method is intrathecal.In some embodiments, administration method is intravenous.In some embodiments In, administration method is subcutaneous.In some embodiments, administration method is intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The horizontal amount of the complement component pyrolysis product of (for example, in the fluid of individual, tissue or organ) in individual.In some cases Under, the theme medicine group of " effective quantity " of the anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure " effective quantity " for closing object is when being applied to individual in need with one or more dosage, keep in individual (for example, individual In fluid, tissue or organ) classic complement approach activity level with not no C1s Antybody therapies anti-with humanization when, such as It is compared and is reduced to the level of the complement component pyrolysis product in fluid, tissue or organ before the anti-C1s Antybody therapies of humanization Few 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount, wherein keeping reducing about 4 hours to about 30 It a period of time is (for example, 4 hours to 8 hours, 8 hours to 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days, 14 days To 21 days or 21 days to 30 days).In some embodiments, individual is mammal.In some embodiments, individual is People.The application of the anti-C1s antibody of humanization can by any approach well known by persons skilled in the art, including it is disclosed herein that A little approach.In some embodiments, administration method is intrathecal.In some embodiments, administration method is intravenous.One In a little embodiments, administration method is subcutaneous.In some embodiments, administration method is intramuscular.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is reduced to when individual application in need with one or more dosage The horizontal amount of the complement component pyrolysis product of (for example, in the fluid of individual, tissue or organ) in individual.In some cases Under, the theme medicine group of " effective quantity " of the anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure " effective quantity " for closing object is when being applied to individual in need with one or more dosage, keep in individual (for example, individual In fluid, tissue or organ) complement component pyrolysis product level with not no C1s Antybody therapies anti-with humanization when, such as It is compared and is reduced to the level of the complement component pyrolysis product in fluid, tissue or organ before the anti-C1s Antybody therapies of humanization Few 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount, wherein keeping reducing about 4 hours to about 21 days a period of time (for example, 4 hours to 8 hours, 8 hours To 24 hours, 2 days to 4 days, 4 days to 7 days, 7 days to 14 days or 14 days to 21 days).In some embodiments, individual is lactation Animal.In some embodiments, individual is people.The application of the anti-C1s antibody of humanization can be by those skilled in the art Any approach known, including those disclosed herein approach.In some embodiments, administration method is intrathecal.In some realities It applies in scheme, administration method is intravenous.In some embodiments, administration method is subcutaneous.In some embodiments, it applies It is intramuscular with approach.

In some cases, " effective quantity " of the anti-C1s antibody of the humanization of the disclosure or the humanization comprising the disclosure resist " effective quantity " of the theme pharmaceutical composition of C1s antibody is made a to when individual application in need with one or more dosage The C4b2a of (for example, in the fluid of individual, tissue or organ) is (that is, Complement C4 b and C2a compound in body；Also referred to as " C3 is converted Enzyme ") generation with not no C1s Antybody therapies anti-with humanization when, such as in individual before the anti-C1s Antybody therapies of humanization or The amount of the C4b2a generated in fluid, tissue or organ compared to reduce at least 10%, at least 20%, at least 30%, at least 40%, At least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% or 100% amount.In some embodiments, individual is mammal.In some embodiments, individual is people.It applies With any approach well known by persons skilled in the art, including those disclosed herein approach can be passed through.In some embodiments In, using being intravenous.In some embodiments, administration method is intrathecal.In some embodiments, administration method is quiet In arteries and veins.In some embodiments, administration method is subcutaneous.In some embodiments, administration method is intramuscular.

Present disclose provides the methods for adjusting complement activation.The method inhibits complement activation in some embodiments, Such as reduce the generation of C4b2a.In some embodiments, present disclose provides adjust disease or illness with complement-mediated Individual in complement activation method, the method includes to the individual apply the disclosure the anti-C1s antibody of humanization or this Disclosed pharmaceutical composition, wherein described pharmaceutical composition include the anti-C1s antibody of humanization of the disclosure.In some embodiments In, such method inhibits complement activation.In some embodiments, individual is mammal.In some embodiments, individual For people.Using any approach well known by persons skilled in the art, including those disclosed herein approach can be passed through.In some realities It applies in scheme, using being intravenous.In some embodiments, using being intrathecal.In some embodiments, using for skin Under.In some embodiments, administration method is intramuscular.

The disease or illness of complement-mediated are the amounts of complement C1s in the cell, tissue, fluid or organ for be characterized in that individual The illness of the horizontal abnormality of exception or complement C1s proteolytic activities.

In some cases, the disease of complement-mediated or illness are characterized in that existing in cell, tissue or fluid and increase The C1s amounts of (higher than normal) or raised complement C1s activity levels.For example, in some cases, the disease or disease of complement-mediated Disease is characterized in that in brain tissue and/or celiolymph, there are raised C1s amounts and/or raised C1s activity.Cell, tissue Or in fluid the C1s amounts of " higher than normal " show the amount of C1s in cell, tissue or fluid higher than normally, control level, such as Higher than the individual of cohort or normal, the control level of groups of individuals.C1s in cell, tissue, organ or fluid " higher than normal " Activity level shows the proteolytic cleavage realized by C1s in cell, tissue, organ or fluid higher than normal, control level, Such as higher than the individual of cohort or normal, the control level of groups of individuals.In some cases, suffer from complement-mediated disease or The individual of illness shows one or more accessory symptoms of such disease or illness.

In other cases, the disease of complement-mediated or illness are characterized in that existing in cell, tissue or fluid and be less than Normal C1s amounts or lower complement C1s activity levels.For example, in some cases, the disease or illness feature of complement-mediated It is that in brain tissue and/or celiolymph, there are lower C1s amounts and/or lower C1s activity.Cell, tissue or fluid In " less than normal " C1s amounts show the amount of C1s in cell, tissue or fluid less than normally, control level, such as less than together The individual of age group or normal, the control level of groups of individuals.C1s activity level tables in cell, tissue or fluid " less than normal " Bright, the proteolytic cleavage realized by C1s in cell, tissue or fluid is less than normal, control level, such as less than cohort Normal, the control level of individual or groups of individuals.In some cases, the individual of the disease with complement-mediated or illness is shown One or more accessory symptoms of such disease or illness.

The disease or illness of complement-mediated are that the wherein amount or activity of complement C1s reaches and cause disease or illness in individual Degree disease or illness.In some embodiments, the disease of complement-mediated or illness are selected from the group being made up of：Together Kind of immunological diseases, autoimmune disease, cancer, blood disease, infectious diseases, inflammatory disease, ischemia reperfusion injury, Neurodegenerative disease, Neurodegenerative conditions, eye disease, nephrosis, graft rejection, vascular diseases and vasculitis diseases.At some In embodiment, the disease or illness of complement-mediated are autoimmune disease.In some embodiments, the disease of complement-mediated Disease or illness are isoimmunization disease.In some embodiments, the disease of complement-mediated or illness are cancer.In some implementations In scheme, the disease or illness of complement-mediated are infectious diseases.In some embodiments, the disease or illness of complement-mediated For inflammatory disease.In some embodiments, the disease of complement-mediated or illness are blood disease.In some embodiments, it mends The disease or illness that body mediates are ischemia reperfusion injury.In some embodiments, the disease or illness of complement-mediated For eye disease.In some embodiments, the disease of complement-mediated or illness are nephrosis.In some embodiments, complement-mediated Disease or illness be graft rejection.In some embodiments, the disease of complement-mediated or illness are antibody-mediated transplanting Repel.In some embodiments, the disease of complement-mediated or illness are vascular diseases.In some embodiments, complement is situated between The disease or illness led are vasculitis illness.In some embodiments, the disease of complement-mediated or illness are nervus retrogression Disease or illness.In some embodiments, the disease of complement-mediated or illness are neurodegenerative disease.In some embodiment party In case, the disease or illness of complement-mediated are Neurodegenerative conditions.

The disease of complement-mediated or the example of illness include but not limited to senile macular degeneration, Alzheimer's disease (Alzheimer ' s disease), amyotrophic lateral sclerosis, allergic reaction, Argyrophilic grain dementia, arthritis are (for example, class wind Wet arthritis), asthma, atherosclerosis, Atypical Hemolytic Uremic Syndrome, autoimmune disease (including example Such as autoimmune hemolytic anemia (AIHA)；Warm antibody AIHA；Mixed type AIHA；Deng), barraquer-Simons syndrome (Barraquer-Simons syndrome), Bei Saiteshi diseases (Disease), Britain's type amyloid angiopathy, big Blister pemphigoid, cypress Graves disease (Buerger ' s disease), C1q nephrosis, cancer, pernicious antiphospholipid syndrome, brain starch Angiopathy, cold coagulation disease, corticobasal degeneration, gram refined Er Shi diseases (Creutzfeldt-Jakob disease), Crow Grace disease (Crohn ' s disease), condensation compositions vasculitis, dementia pugilistica, dementia with Lewy body (DLB), diffusivity god It tangles with calcification, lupus erythematosus discoides, Down syndrome (Down ' s syndrome), Yi Wen Cotards through fibrinogen (Evan ' s syndrome), focal segmental glomerulosclerosis, formal thought disorder, Frontotemporal dementia (FTD), 17 dyes The relevant Frontotemporal dementia of colour solid is with parkinson's syndrome, frontotemporal lobar degeneration, Jie Ciman-Si Tuosile-history mattress kirschner disease (Gerstmann-Straussler-Scheinker disease), actue infectious polyradiculoneuritis (Guillain-Barr é Syndrome), hallervorden-Spatz disease (Hallervorden-Spatz disease), hemolytic uremic syndrome, heredity blood Pipe oedema, hypophosphatasia (hypophosphastasis), idiopathic pneumonia syndrome, immune-complex disease (ICD), inclusion body Myositis, infectious diseases are (for example, by bacterium (for example, Neisseria meningitidis (Neisseria meningitidis) or chain Coccus (Streptococcus)), disease caused by viral (for example, human immunodeficiency virus (HIV)) or other infectors), Inflammatory disease, ischaemic/reperfusion injury, mild cognitive impairment, immune thrombocytopenic purpura (ITP), A type molybdenums it is auxiliary because Son lacks (MoCD), I types membrano proliferative glomerulonephritis (MPGN), II types membrano proliferative glomerulonephritis (MPGN) (fine and close object Storage disorders), membraneous nephritis, multiple infarct dementia, lupus (for example, systemic lupus erythematosus (SLE)), glomerulonephritis, river Rugged disease (Kawasaki disease), multifocal motor neuropathy, multiple sclerosis, multi-system atrophy, myasthenia gravis, cardiac muscle Infraction, myotonia dystrophy, neuromyelitis optica, c-type Niemann-Pick disease (Niemann-Pick disease), with The non-Guamanian motor neuron diseases of neurofibrillary tangles, Parkinson's disease (Parkinson ' s disease), with The Parkinson's disease of dementia, paraoxysmal nocturnal hemoglobinuria, pemphigus vulgaris, Pick's disease (Pick ' s Disease), parkinsonism after encephalitis, polymyositis, prion protein cerebral amyloid angiopathy, progressive subcortical gliosis, Stein-leventhal syndrome, psoriasis, septicemia, shiga toxin Escherichia coli (Shiga-toxin E coli, STEC)-HuS, Spinal muscular atrophy, apoplexy, subacute sclerosing panencephalitis, only entanglement type dementia, graft rejection, vasculitis are (for example, ANCA Associated vasculitis), Wei Genashi granulomas (Wegner ' s granulomatosis), drepanocytosis, cryoglobulin blood Disease, Combination cryoglobulinemia, essential mixed cryoglobulinemia, II type Combination cryoglobulinemia, type III are mixed Close property cryoglobulinemia, ephritis, drug-induced thrombopenia, lupus nephritis, posterior bullous epidermis release, Retardance hemolytic blood transfusion reaction, hypocomplementemia urticarial vasculitis syndrome, paracrystals shape bullous keratopathy become and Inefficacy of Platelets Transfusion (platelet refractoriness).

In some embodiments, the disease of complement-mediated or illness include Alzheimer's disease.In some embodiment party In case, the disease or illness of complement-mediated include Parkinson's disease.In some embodiments, the disease or illness of complement-mediated Including graft rejection.In some embodiments, the disease of complement-mediated or illness are antibody-mediated graft rejection.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure prevents or postpones the disease of complement-mediated in individual The breaking-out of at least one symptom of disease or illness.In some embodiments, individual is mitigated or eliminated in the anti-C1s antibody of the disclosure The disease of middle complement-mediated or at least one symptom of illness.The example of symptom include but not limited to autoimmune type disease, Cancer, blood disease, infectious diseases, inflammatory disease, ischemia reperfusion injury, neurodegenerative disease, nervus retrogression Illness, nephrosis, graft rejection, eye disease, vascular diseases or the relevant symptom of vasculitis illness.Symptom can be neurological symptoms result, For example, cognitive impairment, memory impairment, loss of motor function etc..Symptom can also be in the cell, tissue or fluid of individual C1s protein actives.Symptom can also be the degree of the cell of individual, tissue or the complement activation in fluid.

In some embodiments, the anti-C1s antibody of humanization that the disclosure is applied to individual has adjusted the cell of individual, group Knit or fluid in complement activation.In some embodiments, to individual apply the anti-C1s antibody of theme inhibit individual cell, Complement activation in tissue or fluid.For example, in some embodiments, when the anti-C1s antibody of theme humanization is with one or more When a dosage is administered to the individual of disease or illness with complement-mediated as monotherapy or with combination treatment, and with anti- Complement activation before C1s Antybody therapies in individual is compared, and the complement activation at least about 10%, at least about in the individual is inhibited 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure reduces depositions of the C3 on red blood cell；Example Such as, in some embodiments, the anti-C1s antibody of the disclosure reduces the deposition on RBC such as C3b, iC3b.In some embodiment party In case, the anti-C1s antibody of the disclosure inhibits the hemolysis of complement-mediated.

In some embodiments, the anti-C1s antibody of the humanization of the disclosure reduces depositions of the C3 on blood platelet；For example, In some embodiments, the anti-C1s antibody of the disclosure reduces the deposition on blood platelet such as C3b, iC3b.

In some embodiments, cause selected from the group being made up of using the anti-C1s antibody of the humanization of the disclosure As a result：(a) reduction of complement activation；(b) raising of cognitive function；(c) reduction of neurone loss；(d) Glial Activation Reduction；(e) reduction of lymphocytic infiltration；(f) reduction of macrophages infiltration；(g) reduction of antibody deposition；(h) colloid The reduction of loss cell；(i) reduction of oligodendroglia loss；(j) reduction of dendritic cells infiltration；(k) neutrophil(e) granule is thin The reduction of born of the same parents' infiltration；(l) reduction of hemolysis；(m) reduction of red blood cell phagocytosis；(n) blood platelet phagocytosis is made Reduction；(o) reduction of thrombocytolysis；(p) raising of Graft survival rate；(q) macrophage-mediated phagocytosis Reduction；(r) improvement of eyesight；(s) improvement of motion control；(t) improvement of thrombosis；(u) improvement of blood coagulation；(v) kidney The improvement of function；(w) reduction of antibody-mediated complement activation；(x) reduction for the complement activation that autoantibody mediates；(y) poor The improvement of blood；(aa) reduction of demyelinate；(ab) reduction of eosinophilia；(ac) depositions of the C3 on red blood cell It reduces (for example, reduction of the deposition on RBC such as C3b, iC3b)；Depositions of (ad) C3 on blood platelet reduction (for example, The reduction of the deposition on blood platelet such as C3b, iC3b)；The reduction that (ae) anaphylatoxin generates；(af) autoantibody mediates The reduction that blister is formed；(ag) reduction for the itch that autoantibody induces；(ah) reduction for the lupus erythematosus that autoantibody induces； (ai) reduction for the skin erosion that autoantibody mediates；(aj) reduction that the red blood cell caused by infusion reaction destroys； (ak) reduction of the hemolysis caused by allo-antibody；(al) reduction of the haemolysis caused by infusion reaction； (am) reduction for the thrombocytolysis that allo-antibody mediates；(an) reduction of the thrombocytolysis caused by infusion reaction； (ao) reduction of Mast cell activation；(ap) reduction of mast cell histamine release；(aq) reduction of vascular permeability；(ar) water Swollen reduction；(as) reduction of deposition of the complement on graft endothelium；(at) anaphylatoxin in graft endothelium generates Reduction；(au) reduction for the separation that dermal-epidermal has a common boundary；(av) reduction that the anaphylatoxin during dermal-epidermal has a common boundary generates； (aw) reduction for the complement activation that allo-antibody mediates in graft endothelium；(ax) damage of antibody-mediated myoneural junction The reduction of mistake；(ay) at myoneural junction complement activation reduction；(az) at myoneural junction anaphylatoxin generate subtract It is few；(ba) at myoneural junction complement deposit reduction；(bb) paralysis is reduced；(bc) numb to reduce；(bd) bladder control increases By force；(be) defecation control enhancing；(bf) with the reduction of the relevant death rate of autoantibody；(bg) and the relevant hair of autoantibody The reduction of sick rate.

In some embodiments, when the anti-C1s antibody of theme using one or more dosage as monotherapy or with combination When therapy is administered to the individual of disease or illness with complement-mediated, and with the result in individual before anti-C1s Antybody therapies Horizontal or degree is compared, can realize one or more reductions at least about 10% in following result, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%：(a) complement activation；(b) decrease of cognitive function；(c) neurone loss；(d) colloid is thin Born of the same parents activate；(e) lymphocytic infiltration；(f) macrophages infiltration；(g) antibody deposition；(h) spongiocyte loses；(i) less dash forward glue Cell plastid loses；(j) dendritic cells infiltrate；(k) neutrophil cell infiltrates；(l) hemolysis；(m) red blood cell gulps down The effect of biting；(n) blood platelet phagocytosis；(o) thrombocytolysis；(p) graft rejection；(q) macrophage-mediated phagocytosis is made With；(r) vision loss；(s) antibody-mediated complement activation；(t) complement activation that autoantibody mediates；(u) demyelinate；(v) Eosinophilia.

In some embodiments, when the anti-C1s antibody of theme using one or more dosage as monotherapy or with combination When therapy is administered to the individual of disease or illness with complement-mediated, and with the result in individual before anti-C1s Antybody therapies Horizontal or degree is compared, can realize one or more improvement at least about 10% in following result, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%：A) cognitive function；B) Graft survival rate；C) eyesight；D) motion control；E) thrombus It is formed；F) blood coagulation；G) renal function；And h) hematocrit (erythrocyte counts).

In some embodiments, the benefit in the individual is reduced using the anti-C1s antibody of humanization of the disclosure to individual Body activates.For example, in some embodiments, when the anti-C1s antibody of theme using one or more dosage as monotherapy or with When combination treatment is administered to the individual of disease or illness with complement-mediated, and with the benefit in individual before anti-C1s Antybody therapies Body activation compare, make it is described individual in complement activation reduce at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the cognitive function of the individual is improved using the anti-C1s antibody of the humanization of the disclosure.Example Such as, in some embodiments, it is applied when the anti-C1s antibody of theme using one or more dosage as monotherapy or with combination treatment When with individual to disease or illness with complement-mediated, before with anti-C1s Antybody therapies compared with individual cognitive function, make The cognitive function of the individual improves at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, At least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the cognition work(in the individual is reduced using the anti-C1s antibody of the humanization of the disclosure It can fall off rate.For example, in some embodiments, when the anti-C1s antibody of theme is using one or more dosage as monotherapy Or when being administered to the individual of disease or illness with complement-mediated with combination treatment, and in individual before anti-C1s Antybody therapies Decrease of cognitive function rate compare, make it is described individual in decrease of cognitive function rate reduce at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the anti-C1s antibody of humanization of the disclosure is applied to individual to be reduced in the individual Neurone loss.For example, in some embodiments, when the anti-C1s antibody of theme is using one or more dosage as monotherapy Or when being administered to the individual of disease or illness with complement-mediated with combination treatment, and in individual before anti-C1s Antybody therapies Neurone loss compare, make it is described individual in neurone loss reduce at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the anti-C1s antibody of humanization of the disclosure is applied to individual to be reduced in the individual Glial Activation.For example, in some embodiments, when the anti-C1s antibody of theme is using one or more dosage as single treatment Method or when being administered to the individual of disease or illness with complement-mediated with combination treatment, and with individual before anti-C1s Antybody therapies In Glial Activation compare, make it is described individual in Glial Activation reduce at least about 10%, at least about 15%, at least About 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least About 80%, at least about 90% or be more than 90%.In some embodiments, the spongiocyte is that astrocyte or small colloid are thin Born of the same parents.

In some embodiments, the anti-C1s antibody of humanization of the disclosure is applied to individual to be reduced in the individual Lymphocytic infiltration.For example, in some embodiments, when the anti-C1s antibody of theme is using one or more dosage as single treatment Method or when being administered to the individual of disease or illness with complement-mediated with combination treatment, and with individual before anti-C1s Antybody therapies In lymphocytic infiltration compare, so that the lymphocytic infiltration in the individual is reduced by least about 10%, at least about 15%, at least About 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least About 80%, at least about 90% or be more than 90%.

In some embodiments, the anti-C1s antibody of humanization of the disclosure is applied to individual to be reduced in the individual Macrophages infiltration.For example, in some embodiments, when the anti-C1s antibody of the humanization of the disclosure is with one or more dosage When being administered to the individual of disease or illness with complement-mediated as monotherapy or with combination treatment, and with anti-C1s antibody Macrophages infiltration before treatment in individual is compared, and the macrophages infiltration in the individual is made to be reduced by least about 10%, at least About 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least About 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, the anti-C1s antibody of humanization of the disclosure is applied to individual to be reduced in the individual Antibody deposition.For example, in some embodiments, when the anti-C1s antibody of the humanization of the disclosure using one or more dosage as Monotherapy or when being administered to the individual of disease or illness with complement-mediated with combination treatment, and with anti-C1s Antybody therapies Antibody deposition in preceding individual is compared, so that the antibody deposition in the individual is reduced by least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

In some embodiments, reduce the allergy poison in the individual using the anti-C1s antibody of the disclosure to individual Plain (for example, C3a, C4a, C5a) is generated.For example, in some embodiments, when the anti-C1s antibody of the humanization of the disclosure is with one When a or multiple dosage are administered to the individual of disease or illness with complement-mediated as monotherapy or with combination treatment, with The level generated with the anaphylatoxin in individual before anti-C1s Antybody therapies is compared, and the anaphylatoxin generation in the individual is made to subtract Few at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90% or be more than 90%.

The anti-C1s antibody of humanization present disclose provides the anti-C1s antibody of the humanization of the disclosure or comprising the disclosure and medicine The purposes of the individual of disease or illness of the medicine composite for curing of acceptable excipient with complement-mediated on.At some In embodiment, present disclose provides the humanization of the disclosure anti-C1s Antybody therapies disease or illness with complement-mediated The purposes of individual.In some embodiments, present disclose provides the anti-C1s antibody of the humanization comprising the disclosure and pharmaceutically may be used The purposes of the individual of disease or illness of the medicine composite for curing of the excipient of receiving with complement-mediated.

Present disclose provides the anti-C1s antibody of the humanization of the disclosure production for treat suffer from complement-mediated disease or Purposes in the medicament of the individual of illness.

The anti-C1s antibody of humanization present disclose provides the anti-C1s antibody of the humanization of the disclosure or comprising the disclosure and medicine The pharmaceutical composition of acceptable excipient inhibits the purposes of complement activation on.In some embodiments, the disclosure provides The anti-C1s antibody of humanization of the disclosure or the anti-C1s antibody of humanization comprising the disclosure and pharmaceutically acceptable excipient Pharmaceutical composition inhibit with complement-mediated disease or illness individual in complement activation purposes.In some embodiment party In case, present disclose provides in the individual that the anti-C1s antibody of the humanization of the disclosure inhibits the disease or illness with complement-mediated Complement activation purposes.In some embodiments, the disclosure provides the anti-C1s antibody of humanization and pharmacy for including the disclosure The pharmaceutical composition of upper acceptable excipient inhibits the complement activation in the individual of the disease or illness with complement-mediated Purposes.

Present disclose provides use of the anti-C1s antibody of the humanization of the disclosure in producing the medicament for adjusting complement activation On the way.In some embodiments, the medicament inhibits complement activation.In some embodiments, the medicament inhibits with benefit Complement activation in the individual of disease or illness that body mediates.

Present disclose provides the anti-C1s antibody of humanization for the disclosure in therapeutic treatment or the people sources comprising the disclosure Change the pharmaceutical composition of anti-C1s antibody and pharmaceutically acceptable excipient.In some embodiments, present disclose provides with The anti-C1s antibody of humanization of the disclosure in therapeutic treatment.In some embodiments, present disclose provides controlled for medicine The pharmaceutical composition of the humanization comprising the disclosure anti-C1s antibody and pharmaceutically acceptable excipient in treatment.

Present disclose provides the humanization of the disclosure of the individual for treating disease or illness with complement-mediated is anti- The pharmaceutical composition of C1s antibody or humanization anti-C1s antibody and pharmaceutically acceptable excipient comprising the disclosure.At some In embodiment, present disclose provides the humanizations of the disclosure of the individual for treating disease or illness with complement-mediated Anti- C1s antibody.In some embodiments, present disclose provides the individuals for treating the disease or illness that suffer from complement-mediated The humanization comprising the disclosure anti-C1s antibody and pharmaceutically acceptable excipient pharmaceutical composition.

Present disclose provides the anti-C1s antibody of the humanization of the disclosure for adjusting complement activation or include the people of the disclosure The pharmaceutical composition of sourceization anti-C1s antibody and pharmaceutically acceptable excipient.In some embodiments, present disclose provides The anti-C1s antibody of humanization of the disclosure for adjusting complement activation.In some embodiments, present disclose provides for adjusting Save the pharmaceutical composition of the humanization comprising the disclosure anti-the C1s antibody and pharmaceutically acceptable excipient of complement activation. In some embodiments, the anti-C1s antibody inhibits complement activation.

Embodiment

Following embodiment is proposed to make and use the present invention's to those of ordinary skill in the art's offer Full disclosure and description, and the embodiment is not intended to limit the range for being considered as its invention by inventor, and be not also purport Indicating that experiment below is carried out whole or unique experiment.Made an effort ensure about it is used number (for example, Amount, temperature etc.) accuracy, it is contemplated that arrive some experimental errors and deviation.Unless otherwise stated, otherwise number is parts by weight Number, molecular weight are weight average molecular weight, and temperature is degree Celsius, and pressure is atmospheric pressure or close to atmospheric pressure.The standard of can be used contracting It writes, for example, bp, base-pair；Kb, kilobase；Pl, picoliters；S or sec, second；Min, minute；H or hr, hour；Aa, amino acid； Kb, kilobase；Bp, base-pair；Nt, nucleotide；I.m., intramuscular；I.p., in peritonaeum；S.c., subcutaneously；Etc..

Embodiment 1：Humanization TNT005 variants

Generate the humanization variants of TNT005.Additionally provide the amino acid sequence of the heavy chain VH structural domains of humanization variants 1-5 Row；The nucleotide sequence of the heavy chain VH structural domains of encoding humanized variant.The light of humanization variants 1,2 and 5 is shown in Fig. 6-8 The nucleotide sequence of the amino acid sequence of chain VL structural domains and the light chain VL structural domains of encoding humanized variant.Table 2 and 3 is (respectively For Fig. 9 and 10) amino acid sequence (the VL SEQ ID NO relative to TNT005 are summarized in：7；VH SEQ ID NO：8) ammonia Base acid difference.

Single-letter amino acid code is following (in bracket is three letter amino acid code)：

G- glycine (Gly)

P- proline (Pro)

A- alanine (Ala)

V- valines (Val)

L-Leu (Leu)

I- isoleucines (Ile)

M- methionines (Met)

C- cysteines (Cys)

F- phenylalanines (Phe)

Y- tyrosine (Tyr)

W- tryptophans (Trp)

H- histidines (His)

K- lysines (Lys)

R- arginine (Arg)

Q- glutamine (Gln)

N- asparagines (Asn)

E- glutamic acid (Glu)

D-Asp (Asp)

S- serines (Ser)

T- threonines (Thr)

Embodiment 2：The characterization of humanization TNT005 variants

The binding characteristic of humanization TNT005 variants is provided in table 4 and 5 (being respectively Figure 11 and 12).(the first data of table 4 Row) in provide various humanization TNT005 variants to activate C1s relative binding affinity, be presented in Figure 11.

Generate all 15 kinds of combination (VH variant 1+Vk variants 1；VH variant 1+Vk variants 2；VH variant 1+Vk variants 5；VH becomes Body 2+Vk variants 1；VH variant 2+Vk variants 2；VH variant 2+Vk variants 5；VH variant 3+Vk variants 1；VH variant 3+Vk variants 2； VH variant 3+Vk variants 5；VH variant 4+Vk variants 1；VH variant 4+Vk variants 2；VH variant 4+Vk variants 5；VH variants 5+Vk becomes Body 1；VH variant 5+Vk variants 2；VH variant 5+Vk variants 5；).Each humanization variants is tested to compete with biotinylation TNT005 In conjunction with the ability of active C1s.Data are shown in the second data of Figure 11 row.

Each humanization variants is tested in measuring the commercially available measurement that classical pathway of complement (CP) activates.As a result it is shown in Figure 11 Third data arrange.Data show that all 15 kinds of humanization variants are with the IC similar with TNT005₅₀Inhibit CP activation.

8 kinds of humanization TNT005 variants are combined with the Kinetic Characterization of affinity.Data are depicted in table 5, by it It is presented in Figure 12.

Embodiment 3：The In vivo study of machin

To assess the pharmacokinetics (PK) and pharmacodynamics (PD) property of humanization TNT005, in machin (Macaca inus (Macaca fascicularis)) in carry out single dose and the repeated doses research of humanization TNT005.In addition, to be relatively more logical The bioavailability for crossing the humanization TNT005 of various administration method applies people source by intravascular (IV) or subcutaneous (SC) injection Change TNT005 variants.After humanization TNT005 administrations, take blood plasma and blood serum sample to measure humanization at the specified time point The circulation composition (PK) of TNT005 simultaneously assesses inhibition of the humanization TNT005 to classic complement approach (PD).

All research animals are female, and weight is between 2.4-3.9kg, and between 3-5 Sui.In addition, institute There is animal not receive drug administration.

Research consists of two parts：

1) 1 phase-single dose quantity research compares in the intermedium control (not applying drug), low by intravenous (IV) application The PK/ for matching humanization TNT005 in high dose humanization TNT005 groups applied with subcutaneous (SC) in dosage and high dose group PD；And

2) 2 phases-multi-dose (once a day, continuing 7 days) low dosage SC groups.

1 phase research and design is made of four groups of animals, wherein three groups are administered (n=4 animal/dosage with humanization TNT005 Group), and the 4th group is administered (phosphate buffered saline (PBS) with intermedium control；N=3 animal).IV is administered in peripheral vein Animal provide bolus injection, and the interscapular region at back apply SC injection.It is appointed as control group by the 1st group, and IV is applied Medium.2nd group and the 3rd group is pressed the humanization TNT005 that 10mg/kg and 45mg/kg applies single IV dosage respectively.Finally, 4 groups are applied single SC dosage with the direct SC bioavailabilities compared with corresponding IV groups (the 3rd group) by 45mg/kg.Table 6 summarizes 1 phase research and design.

Table 6

Whole blood is collected respectively in K₂EDTA, which is managed, to be handled in serum separator tube for blood plasma and serum, and is stored immediately At -15 DEG C to -25 DEG C.It is administered with humanization TNT005 or intermedium control in the timetable described according to table 7 in sample collection It is preceding and carry out later.

Table 7

2 phase research and designs are the PK/PD relationships for the repetition low dosage humanization TNT005 for assessing SC applications.Except 1 animal Outside from low dosage IV groups (the 1st group), 2 phase animals are diverted from 1 phase control group (n=3).The animal of 2 phases uses 4mg/kg people daily Source TNT005 SC are administered, and continue 7 days.2 phase animals were administered in 57 days after 1 phase was administered.Table 8 summarizes 2 phase research and designs.

Table 8

Whole blood is collected respectively in K₂EDTA, which is managed, to be handled in serum separator tube for blood plasma and serum, and is stored immediately At -15 DEG C to -25 DEG C.Sample collection carries out before and after the timetable described according to table 9 is administered with humanization TNT005.

Table 9-2 phase whole blood collection timetables

As a result

1 phase pharmacokinetics and pharmacodynamics

To assess the Pharmacokinetic Characteristics of 1 interim humanization TNT005, it is diluted in the blood that specified time point obtains in table 7 Slurry samples simultaneously carry out ELISA to quantify humanization TNT005 plasma concentrations.In short, diluted plasma sample is added in advance It is coated in 96 orifice plates of activation C1s.It is incubated with after subsequent wash in plasma sample, adds the horseradish for having specificity to human IgG The detection antibody of peroxidase conjugated is to detect the humanization TNT005 of C1s combinations.Finally, 3,3 ', 5,5 '-tetramethyls are added Benzidine (TMB) substrate is to cause chrominance response, and comparison colour response is read on spectrophotometer.By from blood plasma sample The standard curve interpolation for the humanization TNT005 that product are run parallel measures humanization TNT005 plasma concentrations for all samples.Figure The pharmacokinetic analysis result of 1 phase of research is shown in 14 (the 1-43 days) and Figure 15 (the 32-43 days).After IV applications, people The blood plasma PK features of source TNT005 show typical higher Cmax, followed by dose-dependent removing.SC applications cause The absorption phase is slow, causes to postpone than overall lower Cmax with matched IV dosage groups faciation.45mg/kg IV and SC dosage groups The clearance rate of middle humanization TNT005 terminates suitable from 72 hours to research.

Figure 14 is depicted in the pharmacokinetics (PK) of humanization TNT005 in the machin of (- the 43 day the 1st day) 1 phase administration Feature.Dosage group receives medium；10mg/kg (MPK) humanization TNT005 IV；45MPK humanization TNT005 IV；Or 45MPK humanization TNT005 SC.The average humanization TNT005 blood plasma that each dosage group is marked and drawed relative to the time after administration is dense It spends (n=4 animal/humanization TNT005 groups).

The pharmacokinetics that Figure 15 is depicted in humanization TNT005 in the machin of (- the 43 day the 32nd day) 1 phase administration is special Sign.Dosage group is identical as Figure 14.The average humanization TNT005 blood plasma that each dosage group is marked and drawed relative to the time after administration is dense It spends (n=4 animal/humanization TNT005 groups).

It usesClassic complement approach kit assesses the pharmacodynamic effects of humanization TNT005.Kit is commercially available, and is related to using enzyme linked immunosorbent assay (ELISA) (ELISA), designed for by work The classical pathway of Activation In Vitro sample and serum sample is estimated in the external generation of final cleavage product (C5b-9) for measuring the approach The active intensity of classic complement approach in product.Illustrate determination sample according to manufacturer.In short, being diluted in the time shown in table 7 The blood serum sample from monkey of point collection is simultaneously added in the hole of 96 orifice plates of offer.After incubation, add to classical pathway Final cleavage product (C5b-9) have specificity detection antibody and chrominance response is measured on spectrophotometer.Compare list The all samples of a monkey and the sample (before administration=100% activity) being standardized as before the administration of identical monkey.Figure 16 shows 1 Phase is grouped the result of pharmacokinetic reading.IV, which applies humanization TNT005, leads to two dosage groups classical pathway upon administration It is immediately exposed to close completely to complete inhibition.Classical pathway it is active recovery be gradually and dose-dependent, 45mg/kg agent The animal ratio 10mg/kg groups for measuring group restore slower.It is shown for pathway activities by IV the and SC groups of 45mg/kg administrations Closely similar recovery time is shown, it is consistent (Figure 14) with the similar humanization TNT005 Pharmacokinetic Characteristics of those groups.

Figure 16 describes the classical pathway activity of the blood serum sample from the machin being administered (- the 43 day the 1st day) 1 phase (PD readings).Dosage group receives medium；10mg/kg (MPK) humanization TNT005 IV；45MPK humanization TNT005 IV；Or 45MPK humanization TNT005 SC.The classical pathway activity that each dosage group is marked and drawed relative to the time after administration (is standardized as Activity before administration；N=4 animal/humanization TNT005 groups).

2 phase pharmacokinetics (PK) and pharmacodynamics (PD)

2 interim humanization TNT005 pharmacokinetics and pharmacodynamics are measured with the same way described with 1 phase.Figure 17 shows The pharmacokinetics and pharmacodynamic analysis result of 2 phases of research are gone out.SC, which applies low dosage humanization TNT005 (4mg/kg), to be caused The slow-absorbing phase (Figure 17, red curve, right side y-axis) in first 24 hours of administration.It is dense with blood plasma humanization TNT005 The raising of degree, serum classical pathway activity reduce (Figure 17, blue curve, left side y-axis).Cause by the daily repeat administrations of 4mg/kg Level before blood plasma humanization TNT005 is gradually increased and is further reduced to administration to the 7th angel's serum classical pathway activity 10% (i.e. -90% classical pathway inhibition).

Figure 17 is depicted in humanization TNT005 in the machin of 2 phases administration (the daily SC applications of 4mg/kg, continue 7 days) PK/PD features.For 2 phases (n=4 animal), average humanization TNT005 plasma concentrations (right side y-axis) are marked and drawed relative to the time It is active (left side y-axis) with average serum classical pathway.

Although having referred to particular embodiment of the invention, invention has been described, those skilled in the art should manage Solution can be variously modified without departing substantially from true spirit and scope of the present invention and available equivalents replace. In addition, can many modifications may be made so that particular condition, material, material composition, technique, one or more steps be suitable for the present invention Purpose, spirit and scope.All such modifications are intended to be within the scope of the following claims.

Claims

1. a kind of humanized antibody of specific binding complement component C1s, wherein the antibody includes：

A) include the areas VH of following amino acid sequence：

(Q/E)VQL(V/Q)QSGAE(V/L)KKPGASVK(L/V)SC(T/A)ASGFNIKDDYIHWV(K/R) QAPGQGLEWIGRIDPADGHTKYAPKFQVK(V/A)TITADTST(S/N)TAY(L/M)(E/Q)LSSL(R/T) SEDTAVYYCARYGYGREVFDYWGQGTTVTVSS(SEQ ID NO：26)；With

B) include the areas VL of following amino acid sequence：

2. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：10 areas VH；With

B) include SEQ ID NO：20 areas VL.

3. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：10 areas VH；With

B) include SEQ ID NO：22 areas VL.

4. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：10 areas VH；With

B) include SEQ ID NO：24 areas VL.

5. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：12 areas VH；With

B) include SEQ ID NO：20 areas VL.

6. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：12 areas VH；With

B) include SEQ ID NO：22 areas VL.

7. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：12 areas VH；With

B) include SEQ ID NO：24 areas VL.

8. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：14 areas VH；With

B) include SEQ ID NO：20 areas VL.

9. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：14 areas VH；With

B) include SEQ ID NO：22 areas VL.

10. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：14 areas VH；With

B) include SEQ ID NO：24 areas VL.

11. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：16 areas VH；With

B) include SEQ ID NO：20 areas VL.

12. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：16 areas VH；With

B) include SEQ ID NO：22 areas VL.

13. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：16 areas VH；With

B) include SEQ ID NO：24 areas VL.

14. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：18 areas VH；With

B) include SEQ ID NO：20 areas VL.

15. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：18 areas VH；With

B) include SEQ ID NO：22 areas VL.

16. humanized antibody according to claim 1, it includes：

A) include SEQ ID NO：18 areas VH；With

B) include SEQ ID NO：24 areas VL.

17. according to the humanized antibody described in any one of claim 1-16, wherein the humanized antibody is selected from by Fab pieces The group of section, 2 segments of F (ab '), scFv and Fv compositions.

18. according to the humanized antibody described in any one of claim 1-16, wherein the humanized antibody includes isotype The heavy chain constant region of IgG1, IgG2, IgG3 or IgG4.

19. a kind of composition, it includes：

A) humanized antibody according to any one of claim 1-18；With

B) pharmaceutically acceptable excipient.

20. composition according to claim 19, wherein the composition includes tonicity agent, suspending agent, emulsifier, stabilization It is one or more in agent, preservative, freeze drying protectant, surfactant and sugar.

21. a kind of container, it includes the compositions according to claim 19 or claim 20.

22. container according to claim 21, wherein the container is sterile.

23. according to the container described in claim 21 or claim 22, wherein the container is bottle, bottle or syringe.

24. a kind of horizontal method reducing complement component pyrolysis product in individual, the method includes pressing effectively to inhibit C1s simultaneously The amount for reducing the pyrolysis product level, to antibody of the individual application according to any one of claim 1-18, or Composition according to claim 19 or 20.

25. according to the method for claim 24, wherein the complement component pyrolysis product is C4 pyrolysis products.

26. according to the method for claim 25, wherein the complement component pyrolysis product is C2 pyrolysis products.

27. according to the method for claim 25, wherein the complement component pyrolysis product is C3 pyrolysis products.

28. according to the method described in any one of claim 24-27, wherein the individual is people.

29. according to the method described in any one of claim 24-28, wherein the application is intravenous.

30. according to the method described in any one of claim 24-28, wherein the application is intramuscular.

31. according to the method described in any one of claim 24-28, wherein the application is intrathecal.

32. according to the method described in any one of claim 24-28, wherein the application is subcutaneous.

33. according to the method described in any one of claim 24-28, wherein described reduce has the illness for treating complement-mediated Effect.

34. according to the method for claim 33, wherein the illness of the complement-mediated is isoimmunization illness.

35. according to the method for claim 33, wherein the illness of the complement-mediated is autoimmune disorder.

36. a kind of method for inhibiting the complement component that C1s is mediated in individual to crack, the method includes pressing effectively C1s to be inhibited to be situated between The amount for the complement component cracking led, to antibody of the individual application according to any one of claim 1-18, or according to Composition described in claim 19 or 20.

37. a kind for the treatment of the method for the disease of complement-mediated or illness in individual, the method includes pressing effectively to treat the benefit The amount of disease or illness that body mediates, to antibody or root of the individual application according to any one of claim 1-18 According to the composition described in claim 19 or 20.